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.

Does the California market squid (Loligo opalescens) spawn naturally during the day or at night? A note on the successful use of ROVs to obtain basic fisheries biology data

The California market squid (Loligo opalescens Berry), also known as the opalescent inshore squid (FAO), plays a central role in the nearshore ecological communities of the west coast of the United States (Morejohn et al., 1978; Hixon, 1983) and it is also a prime focus of California fisheries, ranking first in dollar value and tons landed in recent years (Vojkovich, 1998). The life span of this species is only 7−10 months after hatching, as ascertained by aging statoliths (Butler et al., 1999; Jackson, 1994; Jackson and Domier, 2003) and mariculture trials (Yang, et al., 1986). Thus, annual recruitment is required to sustain the population. The spawning season ranges from April to November and spawning peaks from May to June. In some years there can be a smaller second peak in November. In Monterey Bay, the squids are fished directly on the egg beds, and the consequences of this practice for conservation and fisheries management are unknown but of some concern (Hanlon, 1998). Beginning in April 2000, we began a study of the in situ spawning behavior of L. opalescens in the southern Monterey Bay fishing area.

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.

A sudden collapse in distribution of Pacific sardine (Sardinops sagax) off southwestern Australia enables an objective re-assessment of biomass estimates

Fishery-independent estimates of spawning biomass (BSP) of the Pacific sardine (Sardinops sagax) on the south and lower west coasts of Western Australia (WA) were obtained periodically between 1991 and 1999 by using the daily egg production method (DEPM). Ichthyoplankton data collected during these surveys, specifically the presence or absence of S. sagax eggs, were used to investigate trends in the spawning area of S. sagax within each of four regions. The expectation was that trends in BSP and spawning area were positively related. With the DEPM model, estimates of BSP will change proportionally with spawning area if all other variables remain constant. The proportion of positive stations (PPS), i.e., stations with nonzero egg counts — an objective estimator of spawning area — was high for all south coast regions during the early 1990s (a period when the estimated BSP was also high) and then decreased after the mid-1990s. There was a decrease in PPS from the mid-1990s to 1999. The particularly low estimates in 1999 followed a severe epidemic mass mortality of S. sagax throughout their range across southern Australia. Deviations from the expected relationship between BSP and PPS were used to identify uncertainty around estimates of BSP. Because estimation of spawning area is subject to less sampling bias than estimation of BSP, the deviation in the relation between the two provides an objective basis for adjusting some estimates of the latter. Such an approach is particularly useful for fisheries management purposes when sampling problems are suspected to be present. The analysis of PPS undertaken from the same set of samples from which the DEPM estimate is derived will help provide information for stock assessments and for the management of purse-seine fisheries.

Reproductive biology of the blue swimmer crab (Portunus pelagicus, Decapoda: Portunidae) in five bodies of water on the west coast of Australia

Portunus pelagicus was collected at regular intervals from two marine embayments and two estuaries on the lower west coast of Australia and from a large embayment located approximately 800 km farther north. The samples were used to obtain data on the reproductive biology of this species in three very different environments. Unlike females, the males show a loosening of the attachment of the abdominal flap to the cephalothorax at a prepubertal rather than a pubertal molt. Males become gonadally mature (spermatophores and seminal fluid present in the medial region of the vas deferentia) at a very similar carapace width (CW) to that at which they achieve morphometric maturity, as reflected by a change in the relative size of the largest cheliped. Logistic curves, derived from the prevalence of mature male P. pelagicus, generally had wider confidence limits with morphometric than with gonadal data. This presumably reflects the fact that the morphometric (allometric) method of classifying a male P. pelagicus as mature employs probabilities and is thus indirect, whereas gonadal structure allows a mature male to be readily identified. However, the very close correspondence between the CW50’s derived for P. pelagicus by the two methods implies that either method can be used for management purposes. Portunus pelagicus attained maturity at a significantly greater size in the large embayment than in the four more southern bodies of water, where water temperatures were lower and the densities of crabs and fishing pressure were greater. As a result of the emigration of mature female P. pelagicus from estuaries, the CW50’s derived by using the prevalence of mature females in estuaries represent overestimates for those populations as a whole. Estimates of the number of egg batches produced in a spawning season ranged from one in small crabs to three in large crabs. These data, together with the batch fecundities of different size crabs, indicate that the estimated number of eggs produced by P. pelagicus during the spawning season ranges from about 78,000 in small crabs (CW=80 mm) to about 1,000,000 in large crabs (CW=180 mm).

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.

Larval rearing of Macrobrachium rosenbergii (De Man) in artificial sea water

Larvae of Macrobrachium rosenbergii were successfully reared in artificial sea water prepared in fresh ground water. The water was circulated through a biological filter by means of air-lift pumps for a period of one week to remove the undissolved particles prior to use in the hatchery operation. The experiments were initiated during 1989 and the hatchery has been working on pilot scale since June, 1990. The larvae in all the experiments were fed with egg-custard, Mona and Artemia nauplii. The survival rate varied from 5 to 52% in the 12 experiments. These findings can add to the development of hatcheries in the inland areas which can further boost the popularization of giant freshwater prawn farming.

Observations on the mating behavior, spawn mass and larval development of Hydatina physis Linne, 1758 (Cephalaspidea: Hydatinidae) from Karachi

Reproduction of Hydatina physis was studied in a population from Karachi, Pakistan, including mating and egg laying behavior, spawn characteristics and development.Individuals first appear in the field in October and remain until March. The spawning occurs from mid-November till mid-February with a peak in December. During this period the individuals were also observed pairing. In captivity, mating lasts for 30 minutes, second mating occurs two days later. Oviposition occurs in a very interesting and unusual manner. The mother turns "up-side-down" with its food fully expanded and the shell completely hidden underneath, the expanded foot serves as protective cover to the eggs. Eggs are deposited in a complexly folded mass with a short stem and an adhesive disc. Capsules, arranged in a single layer, contain 4-6 eggs each of wich is 70 um in diameter. Development is planktotrophic and veligers hatch after 14 days at a temperature of 26-28 degrees Celsius.

Rearing of Clarias batrachus (Lin.) larvae with formulated diets

An experiment was carried out for a period of 20 days using 7-day old Clarias batrachus larvae of initial total length of 7.4 ± 0.49 mm and weight of 2.9 ± 0.83 mg. Three artificial diets were used for the study having three replication of each. Among these, diet-I was formulated using 20% fish meal (FM), 30% powdered milk and 30% boiled egg yolk (BEY), diet-II using 27% FM, 20% Baker’s yeast (BY), 30% BEY and 3% agar and diet-III using 20% FM, 20% BY and 45% whole egg. The larvae fed on diets-II and III showed significantly (P<0.05) better length and weight gain than those of the larvae fed on diet-I. The larvae fed on diet- III showed the best survival rate (70%). However, the condition factor of the larvae fed on diet I was significantly better than those of the larvae fed on other two diets. The results of the study showed that C. batrachus larvae could be successfully reared with diet containing 45% whole egg, 20% yeast and 20% fish meal.

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.

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.

Cryopreservation of rhesus macaque (Macaca mulatta) spermatozoa and their functional assessment by in vitro fertilization

Although spermatozoa from several species of nonhuman primates have been cryopreserved, there has been no report of success with rhesus macaque spermatozoa as judged by functional assays. Two Tris-egg yolk freezing media. TEST and TTE. which have: been successfully used for cynomolgus macaque (Macaca fascicularis) spermatozoa, were compared for cryopreservation of spermatozoa From four rhesus macaques (Macaca mulatta). The postthaw motility (percentage and duration) of spermatozoa cryopreserved in TTE was much higher than that for spermatozoa cryopreserved in TEST. The function of sperm cryopreserved in TTE was evaluated by in vitro fertilization or oocytes collected from gonadotropin-stimulated prepubertal rhesus macaques. Of the inseminated oocytes. 82 +/- 13% were fertilized and 63 +/- 22 and 39 +/- 21% of the resulting zygotes developed into morulae and blastocysts. respectively. These results indicate that rhesus macaque spermatozoa can be effectively cryopreserved in TTE medium. This finding will facilitate the application of in vivo and in vitro assisted reproductive technologies in this species. (C) 2001 Academic Press.

Growth and reproductive performance of locally isolated brackish water rotifer (Brachionus plicatilis) feeding different micro algae

Population growth and reproductive capacity of brackishwater rotifer, Brachionus plicatilis, were evaluated, for a period of 8 days in a temperature controlled ( =25°C) microalgallaborarory, under three different algal feeding regimens. The algal species that were tested are: (i) Chlorella sp. (T1), Tetraselmis chui (T2), Nannochloropsis oculata (T 3). The feeding density of each algal species was maintained similar as of 4.5xW6 ceHs mi. The rotifer fed on T. chui showed the highest (p<0.05) population growth (131.5 ind./ml), compared to that fed on Chlorella sp (45.67 ind./ml) and N oculata (43.44 ind./ml). The abundance of egg bearing rotifers was also higher (35.77%) with T. chuithan with Chlorella sp (27.76%) and N oculara (24.60%). The results of the present study indicate that T. chui could be the most suitable algal food for the stock culture of locally isolated rotifer B. plicatilis.