Phenotypic and genetic diversity of coexisting Listonella anguillarum, Vibrio harveyi and Vibrio chagassi recovered from skin haemorrhages of diseased sand smelt, Atherina boyeri, in the Gulf of Trieste (NE Adriatic Sea)

Aims: This study identified and characterized coexisting Vibrios associated with haemorrhagic skin lesion bearing sand smelt fishes (Atherina boyeri) in north‐eastern Adriatic Sea. Methods and Results: Bacteria were isolated from external skin lesions of four samples, and representative morphotypes grown on thiosulfate–citrate–bile salt–sucrose agar were isolated. In total 25 isolates, presumptively assigned to Vibrio genus, were biochemically characterized and were grouped in 10 phenotypic profiles. Phenotypes were heterogeneously distributed among the diseased sand smelt analysed; only one phenotype was recovered from all the samples. Sequencing of 16S rRNA was performed to identify representatives of all phenotypes. Phylogenetic analysis using the neighbour‐joining method revealed six isolates clustered within the Vibrio harveyi group, three clustered with known Vibrio chagasii strains and three clustered with Listonella anguillarum. Conclusions: Vibrios with a broad phenotypic variability were found in the external lesions of diseased A. boyeri. In total three species of Vibrio were identified: V. harveyi showed the wider phenotypical and ribotypical heterogeneity while L. anguillarum shared similar biochemical characteristics with typical strains. Significance and Impact of the study: Previously unreported coexistence of potential pathogenic species colonizing diseased A. boyeri has ecological as well as epidemiological significance.     

Female reproductive biology of the sand smelt in brackish lagoons of southern France

The breeding season of Atherina boyeri living in the brackish lagoons of Mauguio, Pérols and Méjean (Hérault, southern France) was very protracted (February-September). The peak of the reproductive effort occurred in April, May and June. Larger fishes started spawning earlier, and stopped later, than smaller ones. Batch fecundity varied from 4 to 447 oocytes and was positively related to female length and weight. Monthly mean fecundity increased at the beginning of the breeding season and decreased between June and July. Individual mean oocyte diameter ranged from 1.34 to 1.94 mm and was not related to fish length. Monthly mean oocyte size declined throughout the breeding season as water temperature rose. There was no trade-off between number and egg size. The strategy of these sand smelts seemed well adapted to ensure spawning success without endangering their survival. The highest reproductive effort period occurred in such a way that environmental conditions were suitable to the best survival and growth of hatched larvae. When breeding ceased, fishes, particularly the youngest ones, had enough time to improve their condition before overwintering.     

River flow,zooplankton and dominant zooplanktivorous fish dynamics in a warm‐temperate South African estuary

The possible links between river flow, zooplankton abundance and the responses of zooplanktivorous fishes to physico‐chemical and food resource changes are assessed. To this end, the seasonal abundance, distribution and diet of the estuarine round‐herring Gilchristella aestuaria and Cape silverside Atherina breviceps were studied in the Kariega Estuary. Spatio‐temporal differences were determined for selected physico‐chemical variables, zooplankton abundance and zooplanktivorous fish abundance and distribution. Results indicated that, following a river flood event in winter (>30 m³ s^?1), altered physico‐chemical conditions occurred throughout the estuary and depressed zooplankton stocks. Abundance of G. aestuaria was highest in spring, with this species dominant in the upper and middle zones of the estuary, while A. breviceps was dominant in summer and preferred the middle and lower zones. The catch per unit of effort of both zooplanktivores also declined significantly following the flooding, thus suggesting that these fishes are reliant on zooplankton as a primary food source for healthy populations. Copepods dominated the stomach contents of both fish species, indicating a potential for strong interspecific competition for food, particularly in the middle reaches. Temporal differences were evident in dietary overlap between the two zooplanktivorous fish species and were correlated with river flow, zooplankton availability and fish distribution. The findings of this study emphasize the close trophic linkages between zooplankton and zooplanktivorous fishes under changing estuarine environmental conditions, particularly river flow and provide important baseline information for similar studies elsewhere in South Africa and the rest of the world.     

Male reproductive strategy and reserve allocation in sand smelt from brackish lagoons of southern France

In male sand smelt Atherina boyeri , from three coastal brackish lagoons of southern France, the duration of the reproductive season differed according to fish size: larger fish exhibited an extended reproductive season from February to August while smaller males reproduced only from April to June-July. Smaller and larger fish showed differences in their seasonal pattern of reserves. Through the cold season, energy content was exhausted for over-winter maintenance and reproductive needs, and was restored just after fish ceased reproducing. In smaller fish, however, reserves were more rapidly restored than in larger fish, and exhibited a decrease in autumn. This result suggested that small fish invested their energy into growth and winter survival rather than reproduction while older fish invested in reproduction. These results are discussed in terms of life history Strategies.     

An electrophoretic investigation of populations of Atherina boyeri Risso, 1810 and A. presbyter Cuvier, 1829 (Teleostei: Atherinidae): genetic evidence in support of the two species

The data obtained using electrophoresis strongly support the specific status of Atherina boyeri Risso, 1810 and A. presbyfer Cuvier, 1829, and thuscontradict the recently proposedsynonymyof the two species. Four populations of A. boyeri and six populations of A. presbyter were assayed for 11 enzymes and general protein using muscle and liver extracts. Eight of the 11 enzymes were shown to be polymorphic at the 95% level. Sixteen loci, encoding 40 putative alleles were consistently resolved in all 10 populations.
The two species were fixed for different alleles at the EST-3 locus. At the G3PDH locus. with the exception of two heterozygotes, all individuals of each species were also homozygous for different alleles. At the PGM locus the common allele was unique to each species.
The mean Nei's genetic distance ( ), over all loci, calculated between populations of A. boyeri (= 0.10 ± 0.06); between populations of A. presbyter ( = 0.02 0.02) and between populations of A. boyeri and A. presbyter ( = 0.42 0.09) indicated the separateness of the two species.
UPGMA cluster analysis based on genetic distances produced a dendrogram whose principal dichotomy resulted in the formation of two clusters. The ordination of populations in the UPGMA cluster analysis strongly reflected the geographic distribution of populations in both species.     

A study of the population biology of Athevina boyeri Risso, 1810 in Aberthaw Lagoon, on the Bristol Channel, in South Wales

The biology of the sixth population of A. boyeri Risso, 1810 to be recorded outside of the Mediterranean basin is described. The 2 year life cycle was similar to that of other populations of the species that have been investigated. A maximum standard length of 93 mm was recorded for 2-year-old fish. Sexual maturity was attained by females at a length of 39 mm. A temporal lag in ovarian development was noted between the two spawning age groups, with the older age group spawning first. Details are provided of batch spawning in A. boyeriand the difficulties of assessing fecundity in batch spawners is discussed. Discrepancies between direct and indirect methods of assessing batch size indicated that the eggs around the largest mode may not represent the next batch to be spawned. Evidence is presented to suggest that in A. boyeri these eggs represent a ' store' from which batches of eggs are subsequently released.     

A multivariate morphometric investigation of Atherina boyeri Risso, 1810 and A. presbyter Cuvier, 1829 (Teleostei: Atherinidae): morphometric evidence in support of the two species

Two recently developed morphometric methodologies, multiple group principal components analysis (MGPCA) and the box truss method of body form transformation are used to re-address whether Atherina boyeri Risso, 1810 and A. presbyter Cuvier, 1829 are morphometrically distinct. The results revealed that morphometric differences are present between A. boyeri and A. presbyter , consistent with expectations of the existence of two species.
Females of the two species were clearly discriminated into two separate groups because the within-group covariances of several characters within populations of each species were different between species i.e., the covariances were heteroscedastic. The occurrence of heteroscedasticity within the data, although statistically invalidating the results of canonical variates anslysis, indicated there to be a large amount of morphological variation between females of the two species. Males were differentiated as a result of differences in body 'shape', particularly the relationship between head 'shape' and body 'shape'.
Several probable reasons are given for the failure of a previous study to differentiate the two species. The data rejects the synonymy of the two species in support of the continued use of the taxa A. boyeri Risso, 1810 and A. presbyter Cuvier, 1829.     

Isolation, characterization and cross-species testing of microsatellites obtained from a sand smelt (Atherina boyeri) genomic library

This study reports the isolation and characterization of 11 polymorphic microsatellites from a sand smelt (Atherina boyeri) genomic library. Enrichment was performed with di-, tri- and tetranucleotide motifs following the FIASCO procedure (fast isolation by AFLP of sequences containing repeats). All loci were found to be in linkage and in Hardy-Weinberg equilibrium. This represents the first microsatellite isolation for the family Atherinidae and the isolated loci were accordingly tested on four additional species of the family: two recognized (A. presbyter and A. hepsetus) and two proposed ('punctata' and 'non-punctata' forms). Moreover their cross-species suitability on Menidia menidia, belonging to the same order but to the family Atherinopsidae, was also tested.     

Sand smelt (Atherina boyeri) migration within the water system of the Camargue,southern France

In the coastal lagoons of Southern France, sand smelt Atherina boyeri, is known as a sedentary species. However part of the sand smelt population living in the brackish water Vaccarès lagoon in the Camargue annually migrates to fresh waters. The movements of sand smelts within the watersystem of the Camargue were studied from 1986 to 1990. Fish traps and fyke nets were set and catches were standardised to Catch Per Unit Effort (CPUE). A Stepwise Multiple Correlation analysis was carried out to link fish abundance with various environmental factors. Between September and November, sand smelts living in the Vaccarès start to colonize fresh waters (drainage canals and then temporary marshes), until March. Spawning occurs from April to June, both in the shallow branches of the canal and in the marshes. From June onwards adults and young-of-the year leave the fresh waters and return to the Vaccarès lagoon where they remain until the next migration. This autumnal migration to freshwater habitats involves mainly fish less than 3 years old, the oldest individuals remaining all year in the Vaccarès. Water level and photoperiod contribute to 20.7% of the variation of CPUE within the drainage canal, while date, water temperature, and rainfall combined, only account for 2.5% of CPUE variations. Date and photoperiod contribute significantly to explaining the patterns of fish migrations between the canal and the marshes. The density of mature sand smelt in the marshes before spawning, varies between 4 and 4346 fish ha^–1. Fry mortality (against number of eggs spawned) over the first two months of life was assessed to vary between 97.5% and 99.9%. Physiological factors such as osmoregulation linked to water temperature are possibly the ultimate factors explaining these unusual annual migrations of sand smelt in the Camargue.     

Environmental factors controlling the spring immigration of two estuarine fishes Atherina boyeri and Pomatoschistus spp. into a Mediterranean lagoon

Atherina boyeri , a pelagic species, mainly entered a Mediterranean lagoon in the Rhone River Delta, during the day, whatever the flow direction. Flow velocity did not appear to be a significant factor and its swimming abilities seemed to be sufficient to cope with the observed velocity. The combined responses of Pomatoschistus spp., benthic species, to the different environmental factors clearly demonstrated a change in behavioural response during their ontogenic development, i.e. a shift from passive to active or active-passive migration. Young Pomatoschistus spp. (<20 mm L _T) exhibited a behaviour that correspond to the classical selective tidal stream transport of young fishes towards estuarine areas (passive drifting). Large Pomatoschistus spp. were also able to enter the lagoon with the flow, but they remained negatively affected by the flow velocity. The diel cycle appeared to be the main factor controlling the immigration of A. boyeri into the lagoon whereas Pomatoschistus spp. seemed more sensitive to hydrographic conditions prevailing at the connection.