Isolation and characterization of nine polymorphic microsatellite markers in the two‐banded sea bream (Diplodus vulgaris) and cross‐species amplification in the white sea bream (Diplodus sargus) and the saddled bream (Oblada melanura)

We have developed nine new microsatellite markers for the two‐banded sea bream (Diplodus vulgaris) from an enriched genome library protocol. All these loci are polymorphic, with mean allelic diversity of 13 (range 5 –21), and expected and observed heterozygosities from 0.641 to 0.932 and 0.428 to 0.914, respectively. Cross‐species tests in two close‐related species of the genus Diplodus (D. sargus and O. melanura) revealed successful amplifications at 8 out of 9 loci, with mean allele number of 4.75 (range 2–8) and 5.50 (range 3 –10), respectively. These results are consistent with the close phylogenetic relationships between the three species, indicating this set of primers might prove useful for studying the levels of genetic diversity and population differentiation in these three species and in other phylogenetically close species of the genus Diplodus and Sparus.     

Morphometric characterization of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa using computer-assisted spermatozoa analysis (ASMA)

As part of a larger study on sperm quality and cryopreservation methods, the present study characterized the head morphometry of sharpsnout sea bream (Diplodus puntazzo) and gilthead sea bream (Sparus aurata) spermatozoa, using both scanning electron microscopy (SEM) and computer‐assisted morphology analysis (ASMA). The latter method has been used rarely in fish and this is its first application on sharpsnout sea bream and gilthead sea bream spermatozoa. Results obtained using SEM are expensive and time‐consuming, while ASMA provides a faster and automated evaluation of morphometric parameters of spermatozoa head. For sharpsnout sea bream spermatozoa, similar head measurement values were obtained using both ASMA and SEM, having a mean ± standard error length of 2.57 ± 0.01 μm vs 2.54 ± 0.02 μm, width of 2.22 ± 0.02 μm vs 2.26 ± 0.04 μm, surface area of 4.44 ± 0.02 μm² vs 4.50 ± 0.04 μm² and perimeter of 7.70 ± 0.02 μm vs 7.73 ± 0.04 μm using ASMA and SEM, respectively. Although gilthead sea bream spermatozoa were found to be smaller than those of sharpsnout sea bream, spermatozoal head morphometry parameters were also found to be similar regardless of evaluation method, having a mean head length of 1.97 ± 0.01 μm vs 1.94 ± 0.02 μm, head width of 1.80 ± 0.01 μm vs 1.78 ± 0.02 μm, surface area of 3.16 ± 0.03 μm² vs 3.18 ± 0.06 μm² and perimeter of 6.52 ± 0.04 μm vs 6.56 ± 0.08 μm using ASMA and SEM, respectively. The results demonstrate that ASMA can be considered as a reliable technique for spermatozoal morphology analysis, and can be a useful tool for studies on fish spermatozoa, providing quick and objective results.     

Sharpsnout sea bream (Diplodus puntazzo) humoral immune response against the parasite Enteromyxum leei (Myxozoa)

The humoral innate immune response of sharpsnout seabream Diplodus puntazzo against the myxozoan Enteromyxum leei was studied. Enteromyxosis was transmitted by cohabitation and a group of uninfected fish served as control. At 5, 12, 19, 26, 40 and 55 days post-exposure (p.e.), control and recipient fish were sampled to determine the prevalence of infection and some humoral innate immune parameters (antiprotease, antitumoral and peroxidase activities). Prevalence of infection was high from day 12 p.e. and reached 100% at days 40 and 55, when intensity of infection was medium to severe. The antiprotease activity was significantly increased in E. leei-exposed fish with respect to control fish at days 12 and 19 p.e. The serum antitumoral activity was slightly lower in recipient than in control fish at all sampling times, except at 40 days p.e., though no statistically significant differences were observed. Serum peroxidases were higher in all recipient fish than in control ones, with the highest stimulation index at 40 days p.e. Within recipient fish, no differences were detected between sampling times in any of the measured activities. The possible implication of these immune factors in the high susceptibility of D. puntazzo to this enteromyxosis is discussed.     

Characterization of eight microsatellite markers in the white sea bream, Diplodus sargus (Teleostei, Sparidae)

The white sea bream, Diplodus sargus (Teleostei, Sparidae), is a species with a high commercial importance in Mediterranean aquaculture. There is currently little information available about the genetic characteristics of cultured populations. In this survey, we have developed eight polymorphic microsatellites for the white sea bream using an enriched genome library protocol. All of them were polymorphic in the 67 individuals tested, 32 of which were wild specimens, and 35 were individuals from a captive F(1) broodstock. These markers can potentially be useful tools for use in population genetic studies.     

The growth of the common two-banded seabream, Diplodus vulgaris (Teleostei, Sparidae), in Canarian waters, estimated by reading otoliths and by back-calculation

The yearly nature of increment formation in the otoliths of 1–9‐year‐old seabream, Diplodus vulgaris (E. Geoffrey Saint‐Hilaire 1817), from the Canary Islands was validated. The marginal increment method showed that the opaque rings were formed in summer, and the translucent rings in winter. The Brody Proportional Hypothesis and the power length–radius relationship used to back‐calculate the growth trajectories of D. vulgaris showed that this growth model could provide reasonable growth estimates in this species. Growth back‐calculation and growth estimates obtained by direct otolith readings were similar. Data on age and size used to estimate the parameters of the von Bertalanffy growth model for D. vulagris from the Canary Islands showed that males and females had similar growth rates.     

Pea protein concentrate in diets for sharpsnout sea bream (Diplodus puntazzo): effects on growth and health status

Four diets for sharpsnout sea bream juveniles (14 g body weight) with four levels of air-processed pea protein concentrate (PPC) (0, 160, 320 and 487 g/kg diet) were tested in triplicate. The experimental diets were isonitrogenous (43% crude protein) and isolipidic (19% ether extract) and the fish were fed to satiation twice a day. After 125 d, fish growth was diminished by the inclusion of PPC. Feed conversion did not show significant differences in any treatment. Neither the body analyses nor the protein and individual essential amino acid retention efficiencies were affected by the inclusion of PPC in the diet. However, histological gut examinations revealed noticeable differences. Fish fed the diet with the highest inclusion level of PPC presented the longest villous length and the most goblet cells, and the width of the lamina propria increased in the anterior intestine. Although no negative changes in nutritive parameters were detected, these alterations might affect nutrient transport, with negative consequences for fish growth. It was concluded that the PPC in the amounts tested here is an inappropriate substitute for fishmeal in diets for sharpsnout sea bream juveniles.     

Two new species of Siriella (Crustacea--Mysidacea) from the southwest coast of Australia

Two new species of Siriella, S.australiensis sp. nov. and S.spinulasp. nov., are described here. Siriella australiensis is distinguishedby the character of the third and fourth pleopods of the male.Siriella spinula is characterized by the short frontal spineof the labrum and the short linguiform telson showing sexualdimorphism.     

The reproductive biology of the genusBaldellia (Alismataceae)

The reproductive biology of the genusBaldellia was studied under cultivation and in nature.B. ranunculoides subsp.repens is self-incompatible whereasB. ranunculoides subsp.ranunculoides andB. alpestris are self-compatible. Bud-pollination occurs in theB. alpestris. Self-incompatibility correlates with the production of fewer, larger, more showy flowers, higher pollen production (but reduced pollen grain size), fewer carpels and smaller seeds. Resource allocation to sexual reproduction is inversely correlated with the degree of vegetative growth.B. ranunculoides subsp.repens produces more ramets than subsp.ranunculoides, which reproduces mainly by seeds.B. alpestris is intermediate between the two since it is self-compatible, produces fewer, larger seed than subsp.ranunculoides and is also able to propagate by vegetative means.     

The reproductive biology of blackspot sea bream Pagellus bogaraveo in captivity. I. gonadal development, maturation and hermaphroditism

Gonadal development, maturation, spawning and hermaphroditism were investigated in captive blackspot sea bream, Pagellus bogaraveo Brünnich, 1768, during the second, third and fourth years of life. The gonads of 224 fish were examined macroscopically and histologically. Undifferentiated gonads were found in fish smaller than 22.0 cm. Adult fish showed four gonadal differentiations: ovotestes with functional testis and quiescent ovary (Mf), ovotestes with functional ovary and regressed testis (mF), ovotestes with both ovary and testis at a resting stage (mf), and ovaries with no male tissue (F). The overall incidence of gonochoric females F was 41%. Functional males Mf were more frequent in age classes 1+ and 2+, whereas functional females mF predominated in the 3+ age class, above 25.0 cm TL. Histological examination revealed testicular degeneration and atrophy in functional females mF. On the basis of both histological data as well as size and age frequency distribution, it is suggested that the reproductive strategy of P. bogaraveo in captivity is characterized by protandrous hermaphroditism, with a high incidence of female gonochorism. Spawning occurred in March–April, at a size of 28.0 cm and age 3 in males and at 29.5 cm and age 4 in females. The gonosomatic index (GSI) remained constantly low (≤ 0.05) throughout the second and third years of life. A significant increase in GSI was noted in both males and females at the accomplishment of the fourth year of life, coinciding with the spawning season. The results are compared with information available on wild P. bogaraveo and discussed with a view to a possible exploitation of this species in aquaculture, through a reliable control of reproduction.     

The gonadal cycle of captive white bream, Diplodus sargus (L.)

A histological study was carried out to follow gonadal changes in captive white bream during the second and third years of life. All but one individual matured as a male. Spawning was found to occur in winter. Environmental factors are likely to have influenced the reproductive activity of the captive fish.     

Food habits of the shortfin mako, Isurus oxyrinchus, off the southwest coast of Portugal

The shortfin mako, Isurus oxyrinchus, is caught in the eastern North Atlantic as a regular bycatch of the surface-drift longline fishery, mainly directed towards swordfish, Xiphias gladius. Stomachs of 112 shortfin mako sharks, ranging in size from 64 cm to 290 cm fork length, showed teleosts to be the principal component of the diet, occurring in 87% of the stomachs and accounting for over 90% of the contents by weight. Crustaceans and cephalopods were also relatively important in this species’ diet, whereas other elasmobranchs were only present in lower percentages. Meal overlap was observed in half of the sampled sharks. No clear trend of prey size selectivity was found, despite smaller individuals seeming incapable of pursuing larger and faster prey. The retention of small prey was also observed in the diet of all sizes of shark. Seasonality in food habits was in accordance with the current availability of food items. The observed vacuity index of 12% is comparable to foraging ecology studies using gillnetting and appears not to be influenced by baited longline gear. Morphological relationships of the digestive system might add important information to the foraging ecology studies and to ecosystem modelling.     

Sorting grid trials to improve size selectivity of red mullet (Mullus barbatus) and annular sea bream (Diplodus annularis) in Turkish bottom trawl fishery

Sorting grids with two different bar spacings (12 mm and 14 mm) were tested to improve size selectivity of the commercially important fish species, red mullet (Mullus barbatus) and annular sea bream (Diplodus annularis), in Turkish bottom trawl fishery. Fishing trials were carried out with R/V ‘Egesüf’ between April and May 2003 in Izmir Bay, Aegean Sea, using a traditional bottom trawl. Selectivity data were collected by the top cover method and analyzed by means of a logistic equation with the maximum likelihood method. Selectivity parameters for individual hauls were obtained with the software program cc 2000 . Mean selectivity was also estimated and compared using the EC Model software that takes between‐haul variations into account. The codend catch size as an additional explanatory variable was used in the comparison. The Kolmogorov–Smirnov (K–S) test was also applied to detect differences between length‐frequency distributions in the upper and lower bags of the 12 and 14 mm bar spacings for red mullet and annular sea bream. The mean L₅₀ values of red mullet were 8.7 and 10.0 cm with the 12 and 14 mm grids, respectively. The L₅₀ value of 14 mm was comparable with the L₅₀ value of the codend mesh size; however, the 12 mm value was rather low. The mean L₅₀ values of 12 and 14 mm bar spacings were 8.8 and 10.4 cm for annular sea bream, respectively; the value of 14 mm bar spacing was very close to 50% size at sexual maturity of the species (10.5 cm). The K–S test indicated length distributions of red mullet and annular sea bream in the 12 and 14 mm upper and lower bags as significantly different (P < 0.05). These results show that improving the size selectivity in a multi‐species fishery using a single selective device is rather difficult. However, higher size selectivity can be obtained when considering the minimum landing size or the 50% sexual maturity size for a given species.     

Age, growth, maturity, mortality, and yield-per-recruit for annular sea bream (Diplodus annularis L.) from the eastern middle Adriatic Sea

The population dynamics parameters of Diplodus annularis from the eastern middle Adriatic Sea were studied. Total lengths of 1704 specimens ranging from 3.3 to 23.0 cm were obtained from commercial and fishery‐independent catches (2000–2002). The species spawns from April through the end of August, with a peak in May. Overall male to female ratio was 1.00 : 1.05. The species is a rudimentary hermaphrodite in the Adriatic Sea. Total lengths (TL) at 50% maturity were 9.0 cm for males and 10.0 cm for females. These estimated sizes were smaller than the minimum legal landing size (MLS = 15 cm) and greater than the actual minimum landing size (L_c = 7.1 cm) for the bottom trawl net. All specimens sampled were fully mature above 13 cm TL. The oldest individual was 13 years old. Length–weight relationship showed close to isometric growth (b = 3.073). Parameters of the von Bertalanffy growth equation were: L_∞ = 23.95 cm; K = 0.126 per year; t₀ = −1.664 year; r² = 0.896. The low value of total mortality (Z = 0.72) was a consequence of the relatively low rate of natural mortality (M = 0.39) and fishing mortality (F = 0.33). The exploitation ratio was E = 0.46. Estimated parameters and the relative yield‐per‐recruit analysis did not indicate any overexploitation of the species in the study area. However, the estimates include uncertainties and require further confirmation, especially of the natural mortality.     

Demographic and genetic structures of white sea bream populations (Diplodus sargus, Linnaeus, 1758) inside and outside a Mediterranean marine reserve

We studied the white sea bream (Diplodus sargus), a protandrous hermaphroditic fish, in two protected and unprotected areas in southwestern France. We observed a significant difference in the demographic structure between the two areas. Females were present in two different age distributions inside and outside the marine reserve with younger females outside. This suggests plasticity in the age of sexual inversion in the case of an exploited population. Genetic differentiation was weak and apparent at only one locus of 26 surveyed (FST = 0.007, p = 0.04). Our data suggest that gene flow between the two areas is important, or the separation between the two sites is recent. Our data on the white sea bream show that fishes inside and outside the marine reserve are very similar genetically, which means that the 'reserve effect' is truly a demographic one, not the result of genetic differences.     

Distribution and abundance of co-occurring chthamalid barnacles Chthamalus montagui and Chthamalus stellatus (Crustacea,Cirripedia) on the southwest coast of Portugal

Two co-occurring chthamalid barnacle species occur on the rocky shores of continental Portugal: Chthamalus stellatus (Poli) and Chthamalus montaguiSouthward. In the present study, patterns of distribution and abundance (density and percentage cover), both vertically (tidal level) and horizontally (wave action), were studied for each species on headlands (more wave action) and not on headlands (less wave action) and at two spatial scales (shore and site) and three tidal levels (low, mid and upper). Chthamalus montagui was significantly more abundant than C. stellatus in both degrees of wave action and at all tidal levels. Despite this great difference in abundance and considering each species individually, C. montagui was more abundant at mid tide-level, while C. stellatus was more abundant at the low tide-level, and C. montagui was less abundant on headlands, while C. stellatus was more abundant on headlands. Small-scale variability (between sites, tens of meters) of abundance of each species was detected, while differences on a larger scale (between shores, tens of kilometres) were not significant.     

Effects of freshwater availability on the summer distribution of otters Lutra lutra in the southwest coast of Portugal

The availability of freshwater has been suggested to strongly influence the distribution of coastal otters Lutra lutra The test this hypothesis, a study was undertaken on the relationships between otter distribution and freshwater availability in a coast where freshwater is a very scarce resource during the summer In this area otters occur mostly in a few small coastal streams but feed largely in the sea Twentynine streams were surveyed in the summer of 1990, and the presence/absence of otter signs was related to nine habitat variables Freshwater availability was found to be the most important factor influencing the occurrence of otters in summer It is suggested that otters favour large streams with good vegetation cover, for these are the most likely to maintain frestwater during the dry periods The distribution of otter signs m streams was also analysed, and it was found that signs tend to concentrate particularly near the mouth of the streams This habitat analysis has some conservation implications, indicating that the decrease of freshwater availability or quality of streams can make these unsuitable for otters, severely decreasing the areas suitable for the species in the southwest coast of Portugal     

黑鲷的胃排空率

在实验室内,１８℃条件下测量了体重在２０－４０ｇ的黑鲷的胃排空率,并用３种数学模型拟合了黑鲷的排空率,结果表明,３种模型都能很好地拟合实验数据,但通过统计学分析发现不论是用湿重还是用于重表示胃含物,线性模型都拟合得最好。用湿重表示时模型为Ｙ＝１．１４－０．０６ｔ（ｒ＾２＝０．８２,ｐ〈０．００１）;而用干重表示时模型为Ｙ＝１．３６－０．０７Ｔ（ｒ＾２＝０．６２,ｐ〈０．００１）,实验还得出体重２０     

Parasites in the thoracic ganglion of Pachygrapsus marmoratus (Brachyura: Grapsidae) from the coast of Portugal

We examined 149 marbled shore crabs, Pachygrapsus marmoratus, from the coast of Portugal for parasites. In particular, we focused our effort on the crab thoracic ganglion. The thoracic ganglion is the largest concentration of nervous tissue in a crab and thus, parasites associated with this organ are well situated to influence host behavior. We found metacercariae of two microphallid trematode species in the thoracic ganglion. We also found a microsporan and an apicomplexan associated with the thoracic ganglion. Other parasites not associated with the thoracic ganglion included gregarine trophozoites which were present in the digestive diverticulae in some of the crabs and the entoniscid isopod, Grapsion cavolini. Metacercariae of one of the trematodes (probably Microphallus pachygrapsi (Deblock and Prevot)), may influence the mortality of its host.