PERMANENT GENETIC RESOURCES: Isolation and characterization of microsatellite loci in the cichlid fish in Lake Victoria, Haplochromis chilotes

Twelve short tandem repeat markers were successfully isolated from a cichlid, Haplochromis chilotes, in Lake Victoria, and characterized in Haplochromis pyrrhocephalus. The microsatellite regions of these markers were found to have between two and 48 alleles with heterozygosity ranging from 0.07 to 0.97. No loci showed significant departures from the Hardy–Weinberg or linkage equilibrium after the Bonferroni correction (P > 0.05). Cross‐species amplification in other cichlids of Lake Victoria, Haplochromis laparogramma, Lithochromis rubripinnis, L. rufus and Haplochromis sp. ‘rockkribensis’, was successful.     

Characterization of microsatellite markers in the squid,Loligo bleekeri (Cephalopoda: Loliginidae)

Loligo bleekeri has a long spawning season and the size of mature males changes during the season: dimorphic (large/small) early in the spawning season and monomorphic (small) later in the spawning season. To understand how copulatory behaviours relate to the dimorphism, we developed five polymorphic microsatellite loci in L. bleekeri. The level of polymorphism ranged from 10 to 22 alleles with expected heterozygosities ranging from 0.79 to 0.93, suggesting that the novel polymorphic loci should be useful for parentage analysis of L. bleekeri.     

Reproductive biology of the armoured catfish Loricariichthys castaneus (Castelnau, 1855) in Lajes reservoir,southeastern Brazil

Sex ratio, gonadal development, breeding season and fecundity of the armoured catfish Loricariichthys castaneus were described to assess its reproductive strategy in a Brazilian tropical reservoir. In total, 226 specimens (199 females and 27 males) were captured from September 2005 to August 2006 and examined in the laboratory. Females outnumbered males and achieved sizes larger than 330 mm TL. Oocyte development, determined by histological analysis, was asynchronous with oocyte size, ranging from pre‐spawning (27–270 μm) to spawning (243–3460 μm), followed by a sharp decrease in the mean oocyte diameter postspawning (590–730 μm) as the spawning proceeded. Spawning occurred throughout most of the year, peaking in August–September and reaching a low in April–May, according to variations in GSI and frequencies of stages of gonadal development. Batch fecundity ranged from 242 to 833 vitellogenic oocytes (relative fecundity = 2.27 oocytes g^?1), averaging 483 oocytes, and was positively related to gonad weight (P = 0.00003). Oocyte diameters ranged from 0.027 to 5.59 mm, with vitellogenic diameters ranging from 2.08 to 5.59 mm. Continuous development of oocytes throughout the year suggests that L. castaneus presents indeterminate fecundity and is a batch‐spawner. These attributes, associated with parental care and a wide reproductive period, correspond to an equilibrium strategy that has proved to be effective in the Lajes reservoir.     

Atlantic capelin (Mallotus villosus) tetranucleotide microsatellites

Twelve microsatellite loci developed for Atlantic capelin (Mallotus villosus) using magnetic bead hybridization enrichment for tetranucleotide microsatellites revealed five loci composed of single repeat elements and six composed of complex repeats. Forty‐four beach‐spawning females from three different northwestern Atlantic Newfoundland beach‐spawning populations were screened at each locus. Loci were polymorphic (two to 59 alleles per locus) and all but two exhibited high heterozygosity (0.86–1). The loci are considered suitable for addressing questions related to fine‐scale population structure, spawning fidelity and survivorship/kinship issues.     

Genetic and morphometric differences demonstrate fine‐scale population substructure of the yellow perch Perca flavescens: need for redefined management units

Whole‐body morphometrics and 15 nuclear DNA microsatellite loci were analysed for 158 Perca flavescens collected during the spawning season from four spawning locations in central Lake Erie, two along the northern shore and two along the southern shore, to evaluate fine‐scale variation (spanning 17–94 km). Results showed significant morphological and genetic differences among P. flavescens from the four locations. The magnitudes of differences were unrelated to geographic distance, demonstrating spatially heterogeneous levels of genetic divergence. These results linked morphometric and genetic variation, showing a discontinuity of scale between currently defined management units and population structure of P. flavescens in Lake Erie, and support that P. flavescens might exist as one or more metapopulations. Findings demonstrate the value of using complementary techniques for evaluating population structure.     

Development and characterization of novel di‐ and tetranucleotide microsatellite markers in Pacific cod (Gadus macrocephalus)

The Pacific cod (Gadus macrocephalus) supports large commercial fisheries in the northern Pacific Ocean and the Bering Sea. Here we characterize 10 polymorphic microsatellite loci isolated from enriched genomic libraries. Loci were screened on a sample of 96 spawning adults. The average number of alleles per locus was 25.3 (range 12–44), with expected heterozygosities (H_E) ranging from 0.54 to 0.97. No significant deviations of genotypic proportions from Hardy–Weinberg equilibrium or linkage equilibrium were detected. These markers will be used in future studies of population structure and mixed stock analysis of this important gadid species.     

Isolation of microsatellites from two species of scleractinian coral

Microsatellites were isolated from two broadcast‐spawning species of scleractinian coral (Platygyra daedalea and Goniastrea favulus) from Australia's Great Barrier Reef. We found 27 microsatellites across both species, although only five loci were polymorphic in each species. Microsatellite loci displayed a wide range of diversity levels with four to 11 alleles per locus (H_O = 0.26–0.91) in P. daedalea and two to seven alleles per locus (H_O = 0.16–0.96) in G. favulus. Most loci showed departures from Hardy–Weinberg equilibrium which may reflect nonrandom mating but may also be related to difficulties associated with coral DNA.     

Reproduction of the introduced oyster Crassostrea gigas (Bivalvia: Ostreidae) cultured on rafts in Spain

The oyster Crassostrea gigas was introduced in Spain for aquaculture purposes; however, until now, it is not known whether populations are established in the wild, being necessary to define whether this species is spawning and which environmental variables trigger this process. The influence of environmental parameters on the reproduction of C. gigas was evaluated from January 2008 to October 2009 with oysters grown on a raft in the Ría de Arousa (Galicia, NW Spain). Temperature and chlorophyll a are directly correlated to sexual maturation. Oysters can mature at temperatures below 14°C. The temperature necessary for spawning differs between the sexes, requiring a temperature above 15°C for males and 18°C for females. Females had a single massive spawn between June and September, while males had partial spawning from May to December with two peaks, one in May–September and another in October–December, with the second peak more pronounced. The first spawning peak is related to high temperatures and concentrations of chlorophyll a, and the second spawning peak is mainly related to the food availability in the water. The spawning asynchrony may be impeding establishment of wild C. gigas populations in Spain.     

Reproductive biology of albacore Thunnus alalunga

Reproductive variables in albacore Thunnus alalunga were evaluated by gonad histology in samples of 132 males (58–118 cm fork length, L_F) and 112 females (59–101 cm L_F) that were collected from the western North Pacific Ocean from 2001 to 2006. In the sex ratio examination, males greatly outnumbered females in large adult fish (L_F > 100 cm). Thunnus alalunga exhibited a protracted spawning period from March to September in the waters off eastern Taiwan and the Philippines, and the peak spawning activity occurred in March and April. Minimum sizes associated with the classification of mature fish were 78 and 83 cm L_F for males and females, respectively. In addition, the largest L_F of immature fish were 93 cm for males and 94 cm for females. The spawning frequency estimate in April was 1·7 days. Batch‐fecundity estimates of 21 females (89–99 cm L_F) ranged between 0·17 and 1·66 million eggs (mean ±s.d . = 0·94 ± 0·43). The relative fecundity estimates of the 21 females ranged between 9·2 and 92·4 oocytes g^?1 body mass (mean ±s.d . = 50·5 ± 22·8). The results presented in this study provide increased information regarding this species' reproductive‐related characteristics than are currently available in stock status determinations.     

Patterns of Aggression and Operational Sex Ratio Within Alternative Male Phenotypes in Atlantic Salmon

The intensity of male–male aggression during mating is predicted to increase with the operational sex ratio (OSR) (the ratio of sexually active males to females). We observed aggressive behaviour in relation to OSR within alternative phenotypes (large anadromous males and small ‘sneaker’ mature male parr) of Atlantic salmon, Salmo salar, as they competed for access to anadromous females in large outdoor arenas immediately prior to spawning events as well as throughout the spawning season. Anadromous males were primarily aggressive outside of spawning events, and their aggression increased with OSR over a narrow range of 0.08–4. By contrast, mature male parr were aggressive just prior to spawning, with a decrease in aggression over a broad range of OSRs from 1 to 7.4. Similarly, literature data for three other salmonid species indicated a decrease in aggression with increasing OSRs over a range of 1–6.33. These observations suggest that there is considerable variability in competitive behaviour, even within alternative phenotypes. Because our data are derived from repeated observations on the same individuals of different genetic origin, further confirmation of these findings is desirable. Nevertheless, our study underscores the importance of measuring competitive behaviour and OSR at ecologically relevant spatial and temporal scales, both of which may differ between alternative maturation phenotypes.     

Sexuality, sex change and maturation patterns in the yellowfin seabream, Acanthopagrus latus (Teleostei: Sparidae) (Houttuyn, 1782)

Macroscopic and histological studies were carried out to describe the reproductive styles and sex reversal and to follow gonadal changes in captive yellowfin seabream during the second year of life. Four reproductive styles are found in Acanthopagrus latus (Houttuyn, 1782): (i) males and females (gonochorism), (ii) functional males, (iii) transitionals and (iv) functional females. The species is a protandric hermaphrodite and begins life as a functional male with testicular zone undergoing active spermatogenesis, while the ovarian zone is arrested at the primary growth (perinucleolar) phase. Males and females were encountered in virtually all size‐classes. Functional males outnumbered the functional females in all size‐classes in which they were encountered. Sex reversal begins in the transitionals from July to August, after spawning in the functional males, at 14.9–20.2 cm standard length (SL) and, by November, maturation of the ovarian tissue begins. A. latus in cages in Kuwait waters spawns from January to April with a peak in February for males, and a peak in March for females and transitionals. Spawning begins in the 18.3–20.2 cm size‐range fish, peaking in the 20.3–22.2 cm size‐range in both males and females and also in the transitionals, although a few of the latter spawn from 14.9 cm SL. In relation to age, spawning begins at 20 months in males and peaks at 21 months. Females begin to spawn at 21 months with a peak at 22 months, while transitionals generally begin to spawn at 20 months, although a few 14–15‐month‐old sex‐changing individuals were encountered. Temperature, either alone or in combination with other unknown factors, triggers spawning in A. latus.     

Mate availability affects female choice in a fish with paternal care: female counterstrategies against male filial cannibalism

Theory predicts that individuals should adopt counterstrategies against intersexual conflict with their mating partners if the counterstrategies are effective and cost-efficient. In fishes, males with parental care often cannibalize their own offspring, which reduces the female’s fitness and creates intersexual conflicts. Males of the goby Rhinogobius flumineus cannibalize more eggs in the nest when they have access to additional females prior to spawning. Thus, it is predicted that females will strategically avoid spawning with males that have high mate availability. In the present study, we experimentally tested this prediction. When sexual pairs were placed in tanks, most females (control females; 21/22) successfully spawned inside the nest. In contrast, when a gravid female (stimulus female) that was housed in a small transparent cage was shown to the experiment pairs prior to spawning, only about half of the females (experiment females; 16/29) spawned inside the nest; the remaining females released unfertilized eggs outside of the nest. Moreover, experiment females infrequently accepted and followed males into nests, and delayed spawning more often than control females. R. flumineus females prefer males that court frequently. Indeed, experiment females that infrequently received courtship tended to spawn outside of the nest. However, infrequent courtship alone could not explain outside-nest spawning, delay in spawning, or the shorter stay of females in nests. These results imply that the presence of a stimulus female dampens female spawning with males. We suggest that R. flumineus females may strategically reject or hesitate to spawn with males that have high mate availability, and that this spawning avoidance may be a counterstrategy against male filial cannibalism.     

Aggregation and spawning of the humphead wrasse Cheilinus undulatus (Pisces: Labridae): general aspects of spawning behaviour

The humphead wrasse Cheilinus undulatus formed resident spawning aggregations daily after high tide at specific locations along the seaward edge of the Palau barrier reef. The location and extent of one aggregation site remained consistent for 6 years with no physical features distinguishing it from adjacent areas. Spawning was documented most months and probably occurred year round with possible seasonal and lunar variation. Spawning males arrived first at the site, followed by females and potentially small primary males. The aggregation female to male sex ratio was estimated to be between 6:1 and 10:1. A maximum of 15 males and 100–150 females were observed at the site. A male courtship posture with the anal fin pointed, the caudal fin folded down and the dorsal fin folded against the body was maintained while swimming a few metres off the bottom in view of females. When ready to spawn females rose up as the posturing male passed and the pair released gametes in a relatively sedate fashion near the surface along the shelf break. No attempted predation on spawning adults was seen. Egg predation after spawning was uncommon. On days with early to mid‐day high tides the spawning period started 2·0–2·5 h after high tide when the speed of lagoon–ocean tidal currents peaked and lasted c. 1 h. On days with later afternoon high tides, spawning occurred sooner after high tide and before current speeds peaked. Other fishes with planktonic eggs spawned at the site as pairs or small groups in a rough succession after high tide with C. undulatus, the last species to spawn.     

Simultaneous polygyny of the gobiid fish Asterropteryx semipunctata in relation to mate availability

Nesting males of Asterropteryx semipunctata conducted spawning behavior with 2–6 females simultaneously. We carried out field observations on a rocky reef in Kagoshima, Japan, to examine the hypotheses that large males will show multi-female spawning behavior because of their mating advantage, and that simultaneous multi-female spawning will occur when the operational sex ratio (OSR; the ratio of receptive males to receptive females) becomes female-biased. Contrary to our prediction, neither the total number of multi-female spawnings during a spawning season nor mean number of spawning females at a time were correlated with nesting male sizes. This indicates that larger males often did not conduct multi-female spawnings. As predicted, the incidence of multi-female spawning followed the change in the OSR over time—as the OSR in the study area became biased toward females, the incidence of multi-female spawnings gradually increased. Our results suggest that mate availability affects mating patterns in A. semipunctata.     

Interspecific variation in spawning time and male mating tactics of the parrotfishes on a fringing coral reef at Iriomote Island,Okinawa

Spawning time and male mating tactics of parrotfishes (family Scaridae) were investigated on a fringing coral reef at Iriomote Island, Okinawa. Spawning was observed in 14 species, and more frequently in more abundant species such as Chlorurus sordidus, Scarus rivulatus and Chlorurus bowersi. At the reef-edge spawning site, C. bowersi spawned at high tide, C. sordidus spawned both at high tide and in the early morning, whereas Calotomus carolinus and most of the Scarus species such as S. rivulatus spawned only in the early morning, mostly 0630–0830 h. Spawning only in the early morning irrespective of tide phase and moon age has seldom been reported from the scarid species of other localities. It is suggested that spawning in the early morning would be adaptive in species such as S. rivulatus, which migrated considerable distances (ca. 500 m) to the inshore feeding sites, in order to minimize feeding losses due to migration. For male mating tactics, pair spawning by territorial TP (terminal phase) males occurred in all 14 species, and streaking and group spawning by nonterritorial small IP (initial phase) males were seen more frequently in more abundant species. Moreover, group spawning by nonterritorial TP males, which were larger than the IP males but smaller than the territorial TP males, frequently occurred in S. rivulatus. Such mating tactics of TP males have not been reported from Scaridae.     

Reproductive biology of the silver pomfret, Pampus argenteus (Euphrasen), in Kuwait waters

The reproductive activities of the silver pomfret, Pampus argenteus (Euphrasen), in Kuwait waters were investigated from March 1996 to February 1998. Observations on the seasonal distribution of maturity stages and variations in seasonal fluctuations in the gonadosomatic index (GSI) confirmed recent findings that the spawning period begins in May. The species has a prolonged spawning period in the females extending from May to August, whereas the males mature in April and ripe specimens were encountered in monthly samples until September. The recruitment pattern confirmed the one breeding season. There are two spawning peaks, the first in May and the second in August. Variations in GSI relative to fish length indicated that females and males are most fecund at about 24.5–26.4 cm and 20.5–22.4 cm length classes, respectively. The males mature earlier than females, at a minimum size of 12.5–14.4 cm, while the females mature at 20.5–22.4 cm. The oocyte diameter‐frequency distribution suggests a serial rhythm of spawning. Fecundity ranged from 28 965 to 455 661 and correlated positively with: (a) standard length (P < 0.006); (b) ovary weight (P < 0.001); and (c) body weight (P < 0.001), and negatively with egg size (P < 0.003).     

Co-variation of gonadosomatic index and parameters of length–weight relationships of Mediterranean horse mackerel, Trachurus mediterraneus (Steindachner, 1868), in the eastern Adriatic Sea

Seasonal changes in length–weight relationships (LWRs), gonadosomatic index (GSI) and condition factor of Mediterranean horse mackerel, Trachurus mediterraneus (Steindachner, 1868), were studied. Ranging from 14.8 to 39.1 cm total length (TL), a total of 1245 specimens (605 males and 640 females) was collected in the eastern Adriatic Sea. Samples were taken at monthly intervals (January–December 2003) using bottom trawls. The b values of LWRs (W = aL^b) varied during the year with season and condition. Including specimens collected in all seasons, the b values showed that growth of both males and females was positively allometric, i.e. specimens changed shape as they gained weight. The GSI reached maximum values in June and a minimum in December. The condition factor was constant during the pre‐spawning period, lowest immediately after spawning (September), and highest at the end of the recovery phase (December).     

Male Parasitic Spawning in Two Species of Triplefin Blenny (Tripterigiidae): Contrasts in Demography, Behaviour and Gonadal Characteristics

Axoclinus nigricaudus and A. carminalis are blennioid fishes from the Gulf of California, Mexico. Males show alternative mating tactics; territorial individuals attract females while non-territorial individuals parasitically spawn with territorial males' females. Demography and gonadal characteristics were related to the degree of parasitic spawning (sperm competition) within and between species. Males of both species showed bimodal size distributions that largely reflected the two tactics. Territorial males spawned much more frequently than non-territorial males, but parasitic spawning was significant. Non-territorial males were more common and rates of parasitic spawning were much higher in A. nigricaudus indicating that it experienced higher sperm competition. In agreement with sperm competition theory, relative testis weight was greater in A. nigricaudus. In A. nigricaudus, the majority of parasitic matings were by non-territorial males, whereas in A. carminalis, both male-types parasitised frequently. Accordingly, only in A. nigricaudus did non-territorial males have relatively heavier testis than territorial males. In both species the testicular gland, that produces accessory products for sperm transfer, was large in territorial males, but small or undeveloped in non-territorial males suggesting male-types differ in fertilisation mode.     

Seasonal variation in fatty acid composition of silver pomfrets,Pampus argenteus (Euphrasen), in Kuwait waters

Proximate and fatty acid composition of wild silver pomfrets, Pampus argenteus, were studied in Kuwait waters for a period of 1 year (November 2007–October 2008) to see whether there were any seasonal compositional differences between males and females. Ten adults (five males, five females) were sampled each month during (i) Pre‐spawning (March–May), (ii) Spawning (June–August), (iii) Post‐spawning (September–November), and (iv) Winter (December–February). Both sexes had significantly (P < 0.05) higher whole body moisture and lower crude protein and lipid contents in winter compared to the respective males and females sampled in other seasons. However, females had significantly higher (9.1%) lipid content during the pre‐spawning season than females in other seasons (7.0–8.2%). The most abundant fatty acid in whole body lipid in both sexes was C16 followed by C18:1n‐9, which accounted for about 31–35% and 22–24% of total lipids, respectively. Males in the pre‐spawning and spawning seasons had significantly higher total monosaturated fatty acids (MUFA) than males and females in post‐spawning and winter. Males had significantly higher total polyunsaturated fatty acids (PUFA) during post‐spawning seasons than females in pre‐spawning and winter seasons. However, there were no significant differences (P > 0.05) in total saturated fatty acids (SFA), PUFA, EPA (eicosapentaenoic acid), DHA (docosahexaenoic acid) or n‐3/n‐6 ratios between respective males and females in different seasons. Livers in males had significantly (P < 0.05) higher MUFA, SFA, PUFA, EPA and DHA than respective females in all months during the spawning season. Female gonads had significantly (P < 0.05) higher MUFA and PUFA but lower SFA content than males in different months during the spawning season. In contrast to the liver, the gonad DHA content and n‐3/n‐6 ratios in females were significantly higher than in males. The gonads from both sexes contained more than double the amount of EPA present in liver; in the case of DHA this was more than three‐fold higher in female gonads, but not in males. Thus, the presence of higher proportions of PUFA, EPA and DHA in gonads, particularly in eggs of silver pomfret, indicates their need for these fatty acids, which may be used as a guideline for dietary essential n‐3 fatty acid requirements for feed formulation of this species. A higher content of DHA in eggs also indicates the higher requirement for DHA in the broodstock diet of silver pomfret.     

Isolation and characterization of microsatellite loci from the Japanese conger eel Conger myriaster

Five polymorphic microsatellite markers were developed in the Japanese conger eel (Conger myriaster) to understand the recruitment mechanisms and population structure of this species, the spawning sites and migration routes of which are unknown. Three of the loci were highly polymorphic and will serve as powerful tools for detailed population studies of the Japanese conger eel. One of the other two loci showed significant departure from Hardy–Weinberg equilibrium and the other showed low allelic diversity.