Combining stable isotopes and skeletal growth marks to detect habitat shifts in juvenile loggerhead sea turtles Caretta caretta

Understanding the phase and timing of ontogenetic habitat shifts underlies the study of a species’ life history and population dynamics. This information is especially critical to the conservation and management of threatened and endangered species, such as the loggerhead sea turtle Caretta caretta. The early life of loggerheads consists of a terrestrial egg and hatchling stage, a posthatchling and juvenile oceanic, pelagic feeding stage, and a juvenile neritic, primarily benthic feeding stage. In the present study, novel approaches were applied to explore the timing of the loggerhead ontogenetic shift from pelagic to benthic habitats. The most recent years of somatic growth are recorded as annual marks in humerus cross sections. A consistent growth mark pattern in benthic juvenile loggerheads was identified, with narrow growth marks in the interior of the bone transitioning to wider growth marks at the exterior, indicative of a sharp increase in growth rates at the transitional growth mark. This increase in annual growth is hypothesized to correlate with the ontogenetic shift from pelagic to benthic habitats. Stable isotopes of carbon and nitrogen just interior and exterior to the transitional growth mark, as well as stable isotopes from pelagic and benthic flora, fauna and loggerhead stomach contents, were analyzed to determine whether this transition related to a diet shift. The results clearly indicate that a dietary shift from oceanic/pelagic to neritic/benthic feeding corresponds to a transitional growth mark. The combination of stable isotope analysis with skeletochronology can elucidate the ecology of cryptic life history stages during loggerhead ontogeny.

Sex-specific growth and mortality, spawning season, and female maturation of the stripey bass (Lutjanus carponotatus) on the Great Barrrier Reef

Sex-specific demography and reproductive biology of stripey bass (Lutjanus carponotatus) (also known as Spanish flag snapper, FAO) were examined at the Palm and Lizard island groups, Great Barrier Reef (GBR).Total mortality rates were similar between the sexes. Males had larger L∞ at both island groups and Lizard Island group fish had larger overall L∞. Female:male sex ratios were 1.3 and 1.1 at the Palm and Lizard island groups, respectively. The former is statistically different from 1, but is unlikely significantly different in a biological sense. Females matured on average at 2 years of age and 190 mm fork length at both locations. Female gonadal lipid body indices peaked from August through October, preceding peak gonadosomatic indices in October, November, and December that were twice as great as in any other month. However, ovarian staging revealed 50% or more ovaries were ripe from September through February, suggesting a more protracted spawning season and highlighting the different interpretations that can arise between gonad weight and gonad staging methods. Gonadosomatic index increases slightly with body size and larger fish have a longer average spawning season, which suggests that larger fish produce greater relative reproductive output. Lizard Island group females had ovaries nearly twice as large as Palm Island group females at a given body size. However, it is unclear whether this reflects spatial differences akin to those observed in growth or effects of sampling Lizard Island group fish closer to their date of spawning. These results support an existing 250 mm minimum size limit for L. carponotatus on the GBR, as well as the timing of a proposed October through December spawning closure for the fishery. The results also caution against assessing reef-fish stocks without reference to sex-, size-, and location-specific biological traits.

Testicular development in migrant and spawning bluefin tuna (Thunnus thynnus (L.)) from the eastern Atlantic and Mediterranean

Testis histological structure was studied in bluefin tuna (Thunnus thynnus) from the eastern Atlantic and Mediterranean during the reproductive season (from late April to early June). Testicular maturation was investigated by comparing samples from bluefin tuna caught on their eastward reproductive migration off Barbate (Strait of Gibraltar area) with samples of bluefin tuna fished in spawning grounds around the Balearic Islands. Histological evaluations of cross sections showed that the testis consists of two structurally different regions, an outer proliferative region where germ cells develop synchronously in cysts, and a central region made up of a well-developed system of ducts that convey the spermatozoa produced in the proliferative region to the main sperm duct. Ultrastructural features of the different stages of the male germ cell line are very similar to those described in other teleost species. The bluefin tuna testis is of the unrestricted spermatogonial testicular type, where primary spermatogonia are present all along the germinative portion of the lobules. All stages of spermatogenesis were present in the gonad tissue of migrant and spawning bluefin tuna, although spermatids were more abundant in spawning fish. The testis size was found to increase by a factor of four (on average) during migration to the Mediterranean spawning grounds, whereas the fat bodies (mesenteric lipid stores associated with the gonads) became reduced to half their weight, and the liver mass did not change significantly with sexual maturation. Linear regression analysis of the pooled data of migrant and spawning bluefin tuna revealed a significant negative correlation between the gonad index (IG) and the fat tissue index (IF), and a weaker positive correlation between the gonad index (IG) and the liver index (IL). Our analyses indicate that the liver does not play a significant role in the storage of lipids and that mesenteric lipid reserves constitute an important energy source for gametogenesis in bluefin tuna.

Maturity, ovarian cycle, fecundity, and age-specific parturition of black rockfish (Sebastes melanops)

From 1995 to 1998, we collected female black rockfish (Sebastes melanops) off Oregon in order to describe their basic reproductive life history and determine age-specific fecundity and temporal patterns in parturition. Female black rockfish had a 50% probability of being mature at 394 mm fork length and 7.5 years-of-age. The proportion of mature fish age 10 or older significantly decreased each year of this study, from 0.511 in 1996 to 0.145 in 1998. Parturition occurred between mid-January and mid-March, and peaked in February. We observed a trend of older females extruding larvae earlier in the spawning season and of younger fish primarily responsible for larval production during the later part of the season. There were differences in absolute fecundity at age between female black rockfish with prefertilization oocytes and female black rockfish with fertilized eggs; fertilized-egg fecundity estimates were considered superior. The likelihood of yolked oocytes reaching the developing embryo stage increased with maternal age. Absolute fecundity estimates (based on fertilized eggs) ranged from 299,302 embryos for a 6-year-old female to 948,152 embryos for a 16-year-old female. Relative fecundity (based on fertilized eggs) increased with age from 374 eggs/g for fish age 6 to 549 eggs/g for fish age 16.

Multidirectional movements of sportfish species between an estuarine no-take zone and surrounding waters of the Indian River Lagoon, Florida

We examined movement patterns of sportfish that were tagged in the northern Indian River Lagoon, Florida, between 1990 and 1999 to assess the degree of fish exchange between an estuarine no-take zone (NTZ) and surrounding waters. The tagged f ish were from seven species: red drum (Sciaenops ocellatus); black drum (Pogonias cromis); sheepshead (Archosargus probatocephalus); common snook (Centropomus undecimalis); spotted seatrout (Cynoscion nebulosus); bull shark (Carcharhinus leucas); and crevalle jack (Caranx hippos). A total of 403 tagged fish were recaptured during the study period, including 65 individuals that emigrated from the NTZ and 16 individuals that immigrated into the NTZ from surrounding waters of the lagoon. Migration distances between the original tagging location and the sites where emigrating fish were recaptured were from 0 to 150 km, and these migration distances appeared to be influenced by the proximity of the NTZ to spawning areas or other habitats that are important to specific life-history stages of individual species. Fish that immigrated into the NTZ moved distances ranging from approximately 10 to 75 km. Recapture rates for sportfish species that migrated across the NTZ boundary suggested that more individuals may move into the protected habitats than move out. These data demonstrated that although this estuarine no-take reserve can protect species from fishing, it may also serve to extract exploitable individuals from surrounding fisheries; therefore, if the no-take reserve does function to replenish surrounding fisheries, then increased egg production and larval export may be more important mechanisms of replenishment than the spillover of excess adults from the reserve into fishable areas.

Seasonal egg production of whitemouth croaker (Micropogonias furnieri) in the Río de la Plata estuary, Argentina-Uruguay

The reproductive biology of the whitemouth croaker (Micropogonias furnieri) inhabiting the estuarine waters of the Río de la Plata (Argentina-Uruguay) was studied by using histological analysis of the ovaries. Samples were collected during the spawning peak and the end of two breeding seasons (November 1995–Feb-ruary 1996 and November 1997–March 1998). Micropogonias furnieri is a multiple spawner with indeterminate annual fecundity. Spawning frequency, determined by using the percentage of females with postovulatory follicles, was about 31% in November 1995 and 25% in February 1996. At these frequencies, a female on average spawned a new batch of eggs every 3–4 days during the spawning season. Batch fecundity was fitted to a power function of length and a linear function of ovary-free female weight. The number of hydrated oocytes decreased at the end of the breeding season, coinciding with an increase of atresia. Annual egg production for a 40-cm-TL female was estimated to be between 3,300,000 and 7,300,000 eggs. In addition to the seasonal decrease in fecundity and spawning activity, a decline in egg size and weight toward the end of the breeding season was also observed.

Assessment of sampling methods to estimate horseshoe crab (Limulus polyphemus L.) egg density in Delaware Bay

Each spring horseshoe crabs (Limulus polyphemus L.) emerge from Delaware Bay to spawn and deposit their eggs on the foreshore of sandy beaches (Shuster and Botton, 1985; Smith et al., 2002a). From mid-May to early June, migratory shorebirds stopover in Delaware Bay and forage heavily on horseshoe crab eggs that have been transported up onto the beach (Botton et al., 1994; Burger et al., 1997; Tsipoura and Burger, 1999). Thus, estimating the quantity of horseshoe crab eggs in Delaware Bay beaches can be useful for monitoring spawning activity and assessing the amount of forage available to migratory shorebirds.

Reproduction and recruitment of white mullet (Mugil curema) to a tropical lagoon (Margarita Island, Venezuela) as revealed by otolith microstructure

The reproductive activity and recruitment of white mullet (Mugil curema) was determined by observations of gonad development and coastal juvenile abundance from March 1992 to July 1993. Adults were collected from commercial catches at three sites in northeastern Venezuelan waters. Spawning time was determined from the observation of macroscopic gonadal stages. Coastal recruitment was determined from fish samples collected biweekly by seining in La Restinga Lagoon, Margarita Island, Venezuela. The examination of daily growth rings on the otoliths of coastal recruits was used to determine their birth date and estimate the period of successful spawning. Fish with mature gonads were present throughout the year but were less frequent between September and January when spawning individuals migrated offshore. In both years, juvenile recruitment to the lagoon was highest between March and June when high densities of 25–35 mm juveniles were observed. Back-calculated hatching-date frequency distributions revealed maximum levels of successful spawning in December–January that were significantly correlated with periods of enhanced upwelling. The relation between the timing of successful spawning and the intensity of coastal recruitment in white mullet was likely due to variations in food availability for first-feeding larvae as well as to variations in the duration of the transport of larvae shoreward as a result of varying current conditions associated with upwelling.

Age, growth, and reproduction of permit (Trachinotus falcatus) in Florida waters

We examined 536 permit (Trachinotus falcatus, 65–916 mm FL) collected from the waters of Florida Keys and from the Tampa Bay area on Florida’s Gulf coast to describe their growth and reproduction.Among permit that we sexed, females ranged from 266 to 916 mm in length (mean=617) and males ranged from 274 to 855 mm (mean=601). Ages of 297 permit ranging from 102 to 900 mm FL were estimated from thin-sectioned otoliths (sagittae). The large proportion of otoliths with an annulus on the margin and an otolith from an OTC-injected fish suggested that a single annulus was formed each year during late spring or early summer.Permit reach a maximum age of at least 23 years.Permit grew rapidly until an age of about five years, and then growth slowed considerably. Male and female von Bertalanffy growth models were not significantly different, and the sexes-combined growth model was FL=753.1(1–e –0.348(Age+0.585)). Gonad development was seasonal, and spawning occurred during late spring and summer over artificial and natural reefs at depths of 10–30 m. Ovaries that contained oocytes in the final stages of oocyte maturation or postovulatory follicles were found during May–July. We estimated that 50% of the females in the population had reached sexual maturity by 547 mm and an age of 3.1 years and that 50% of the males in the population had reached sexual maturity by 486 mm and an age of 2.3 years. Because Florida regulations restrict the maximum size of permit caught in recreational and commercial fisheries to 20-inch (508-mm), most fish harvested are sexually immature. With the current size selectivity of the fishery, the spawning stock biomass of permit could decrease quickly in response to moderate levels of fishing mortality; thus, the regulations in place in Florida to restrict harvest levels appear to be justified.

A comparative study on the hatching of common carp eggs in hapa and hatchery (model CIFE D-80)

Common carp (Cyprinus carpio) eggs were incubated to study the efficiency of hatching in hapa and hatchery. During incubation the recorded temperature was 21-28 degree C and 20-31 degree C, dissolved oxygen 6-9 ppm. and 3-5 ppm., total alkalinity 180-250 ppm. and 28-62 ppm. respectively in the hatchery (model C.I.F.E. D-80) and hapa. CO sub(2) was totally absent in the hatchery, but recorded 3-10 ppm. in the hapa. The flow of water was maintained at 1.25 l/minute/jar in the hatchery. Under the above environmental conditions the eggs hatched in 42-51 hrs. in the hatchery and 61-81 hrs. in the hapa from egg to spawn thereby establishing the hatchery to be a better hatching system for carp eggs.

Studies on the gonadosomatic index and fecundity of chapila (Gudusia chapra Ham.)

The study was conducted in pond to determine the fecundity and gonadosomatic index of chapila, Gudusia chapra. The male fish was found to attain sexual maturity at 7.7 cm and 7.41 g and that of the female at 9.3 cm and 14.65 g by standard-length and body-weight respectively. The investigated fishes were found to be male and female at the ratio of 1:3 and generally female was found to be larger than male. The fish was found to spawn for several months with two spawning peaks, one in April and another in August as indicated by the peaks of gonadosomatic index and ova diameter. Fecundity of the fish ranged from 25,220 to 154,528 with an average value of 72,383 and was found to increase with the increase in length and weight of the fish. The relationships between fecundity and standard-length, body-weight, gonad-length and gonad weight of the fish were found to be linear and significant.

Hatching and rearing of carp (Cyprinus carpio L.) in the Polonnaruwa nursery

Carp fingerlings have been raised at Polonnaruwa since 1957 (Ling, 1962), by a method essentially the same as that described by Hora and Pillay (1962). The present work was initiated to assess and increase the efficiency of the nursery. Two experiments were carried out. In the first, 3 females and 6 males were used. Thirty bundles of Hydrilla were tied to the 3 strings and 10 of them taken at random were used for egg counts. In the second experiment the same number of fish was used but 36 bundles of Hydrilla were tied to the 3 strings and 9 of these taken at random for egg counts. The results of these 2 experiments are given in Table l.

Results of a random stratified bottom trawl survey for scad and mackerel stocks in Mozambican waters: small pelagic fish December-January 1985

The results obtained during the third phase of Nauka are reported concerning the standing stock estimates, population length structure and gonad development of scad and mackerel stocks and the catch composition in Mozambican waters.

Cryopreservation of sperm of common carp, Cyprinus carpio and silver barb, Barbonymus gonionotus for genetically improved seed production

Experiments were conducted to develop and standardize the protocols for cryopreservation of sperm of common carp, Cyprinus carpio and also for using the cryopreserved sperm for fertilization of eggs. Nine extender solutions as Alsever's solution, kurokura-1, kurokura-2, urea egg-yolk, egg-yolk citrate, 0.6% glucose, 0.9% NaCl, Ma and Mb, and five cryoprotectants namely ethanol, methanol, dimethylsulfoxide (DMSO), dimethylamine (DMA) and glycerol were tested. The cryoprotectants were mixed at 10% concentration of the extenders (v/v) to make the cryodiluents. Milt and cryodiluents were mixed at a ratio of 1:9 for Alsever's solution, kurokura-1, kurokura-2, 0.6% glucose and 0.9% NaCl, 1:4 for urea egg-yolk, egg-yolk citrate, Ma and Mb. Among the cryodiluents Alsever's solution mixed with either ethanol or methanol was found to be suitable and it produced more than 90% and 80% spermatozoan motility at equilibrium and post-thaw periods, respectively. Kurokura-1 and kurokura-2 when mixed with the same cryoprotectants showed good spermatozoan motility at equilibrium period (80-90%) but the motility was reduced (30-55%) at post-thaw state. Other extenders did not produce acceptable sperm-motility and in some cases the frozen milt became clotted. Different dilution ratios (1:1, 1:2, 1:4, 1:5, 1:7, 1:9, 1:12, 1:15, 1:20) were formulated for obtaining a suitable milt dilution, the dilution ratio of 1: 9 (milt : cryodiluent) demonstrated the highest post-thaw spermatozoan motility (80%) in Alserver's solution. The optimum concentration of cryoprotectants in the cryodiluents was determined, 10% concentration level was found to be effective to produce the highest number of spermatozoan motility in comparison to the other concentrations (5%, 15%, 20% 30%). Sperm preserved with the cryodiluent Alsever's solution along with either methanol or ethanol was found to be effective to fertilize eggs and produce hatchlings. The hatching rates ranged between 1.48% and 14.76%, compare to control. The fish produced through use of cryopreserved sperm and normal sperm were found to grow well and no significant (P<0.05) growth difference was observed between them. In case of silver barb, Barbonymus gonionotus, sperm tested against six extenders such as egg-yolk citrate, urea-egg-yolk, kurokura-1, kurokura-2, 0.9% NaCl and modified fish ringer (MFR) solution. Cryoprotectants used were the same as those of C. carpio. Milt was diluted with the cryodiluent at a ratio of 1:4 for egg-yolk citrate and urea-egg-yolk, 1:5 for kurokura-1 and 1:9 for 0.9% NaCl, MFR and kurokura-2. The cryoprotectant concentration was maintained at 10% of the extender (v/v) in all the cases. Among the extenders, egg-yolk citrate and urea-egg-yolk mixed with 10% DMSO, methanol and ethanol produced 50% post-thaw spermatozoan motility, whereas DMA and glycerol provided only 10% motility. Trials on milt dilution ratio and cryoprotectant concentration are being conducted. Fertilization trials are also underway.

Sperm cryopreservation of Indian major carp, Labeo rohita: cryodiluents, sperm: cryodiluent dilution ratio and cryoprotectan t concentration

Cryogenic preservation trials of spermatozoa of Labeo rohita were carried out. Twenty four cryodiluents (extender + cryoprotectant), with the combination of six extenders such as egg-yolk citrate, urea-egg-yolk, 0.9% NaCl, Kurokura-2, Ma and Mb and four cryoprotectants viz. DMSO, glycerol, methanol and ethanol, were used to screen out the suitable cryodiluents. Sperm was preserved in 0.25ml plastic straw in programmable freezer. Two step freezing method was followed. Sperm preserved with egg-yolk citrate and urea-egg-yolk containing 10% DMSO showed best post-thaw motility (80%) followed by 0.9% NaCl (60%) and Kurokura-2(30%) solutions. Sperm with the extenders M" and Mb clotted at the time of equilibration and also after few days of preservation. Egg-yolk citrate mixed with ethanol and methanol also showed good percentage of motility (80%) but egg-yolk citrate with glycerol showed less sperm motility (>60%). To determine suitable dilution ratio of milt and cryodiluent two best extender eggyolk citrate and urea-egg-yolk with four cryoprotectants such as DMSO, glycerol, methanol and ethanol at different ratio viz 1:2,1:4,1:7,1:10,1:15 and 1:20 were used. Highest post-thaw motility (>80%) was observed when milt was preserved with egg-yolk citrate containing 10% DMSO at 1:2, 1:4, 1:7 and 1:10 dilutions. Meanwhile using glycerol as cryoprotectants provided less post thaw motility at lower dilution ratio but with the increase of its dilution showed good sperm motility compared with other cryoprotectants. Finally, evaluation on the effect of cryoprotectant concentration on post-thaw sperm motility was conducted. Egg-yolk citrate and four cryoprotectant i.e. DMSO, glycerol, methanol and ethanol with six different concentrations namely 5%,7%, 10%, 15%, 20% and 30%.were evaluated. Among the cryoprotectants DMSO, methanol and ethanol showed highest post-thaw motility (about 80%) at 7% and 10% concentrations. Although glycerol was not suitable at low concentration but its 20% and 30% concentration levels provided best post-thaw motility. No post-thaw motility was obtained with DMSO at 30% concentration. The overall analysis on cryoprotectant concentration indicated that below 5% and above 20% cryoprotectant concentrations could not be suitable for effective cryopreservation of spermatozoa.