Importance of maternal size on the reproductive success of perch,Perca fluviatilis,in small forest lakes: implications for fisheries management

Abstract Abundance, sex ratio and size structure of perch, Perca fluviatilis L., populations were studied in two forest lakes. The influence of maternal size on the amount and quality of eggs and larvae was investigated, and the effect of the spawning stock’s length structure on population egg and fry production was evaluated. The importance of large females was essential for the reproduction of perch. Large perch spawned more and bigger eggs that had higher hatching percentage resulting in considerably higher fry production. Larvae of large females were heavier and survived longer with bare yolk sac reserves. The relation between egg dry weight and female length differed between the study lakes suggesting varying maternal influence in different environments. Decrease in the share of large individuals in a perch population can substantially reduce the amount and quality of larvae produced. The results can be applied to improve the sustainability of perch fisheries.     

Effects of different step‐wise temperature increment regimes during egg incubation of Atlantic cod (Gadus morhua L.) on egg viability and newly hatched larval quality

We carried out an experiment to determine how rapidly the early incubation temperature of Atlantic cod eggs can be increased without affecting normal embryonic development and hatching. Atlantic cod eggs were incubated at a constant low temperature (4.5 ± 0.5°C; T5 – control) and four temperature increment treatments where the temperatures were increased stepwise from 4.5°C at zygote stage to 9.5 ± 05°C (T1‐8 h, T2‐32 h, T3‐64 h and T4‐96 h). Embryonic cell symmetry, embryonic mortality, hatching success and larval skeletal abnormalities, length and yolk sac volume were recorded. Larval samples were also taken at hatch for histological analysis. Except for higher egg mortality and lower hatching success in the T1, the differences among experimental groups were minor. Cell asymmetries and embryo mortalities were not significantly different between the control and T2–T4 treatment groups. Control larvae were significantly longer and had smaller yolk reserves at hatch than T1–T4 larvae and larvae from T2 had the largest yolk reserves. Tissue and organ histology of hatched larvae were similar. Considering embryonic cleavage pattern, hatching success and larval morphology and histology, a gradual increment of temperature in 32 h seems to be the better choice for future developmental programming studies in Atlantic cod.     

Feeding early larval stages of fire shrimp Lysmata debelius (Caridea, Hippolytidae)

An important constraint to the commercial rearing of the marine ornamental shrimp Lysmata debelius is high larval mortality during early stages due to inappropriate procedures of larval collection and not feeding a live prey before one day elapsed after hatching. This incorrect feeding practice is commonly adopted in larval rearing of L. debelius and other ornamental marine shrimps because it is wrongly assumed that reserves of the newly hatched are enough for the first 24 h of life. Present work demonstrates that captive newly hatched L. debelius larvae ingest microalgae within minutes after hatching. When fed solely with Artemia nauplii, they have acceptable survival rates with stocking densities at or below 50 larval L^–1; but when nauplii are combined with microalgae, survival is further improved to zoea 2 as initial mortality is reduced, and higher stocking densities are supported (up to 75 larvae L^–1). The microalgae used were Rhinomonas reticulata, Skeletonema costata and Tetraselmis chuii. Higher survival through metamorphosis to zoea 2 was always observed for groups fed combinations of microalgae including Tetraselmis chuii. It is recommended that, larval collection methods ensure that larvae are fed microalgae within 2–3 h of release.     

Dehydration is more effective for the control of embryonic development and larval hatching of Diplectanum aequans (Monogenea, Diplectanidae) than formalin and trichlorphon

Embryonic development and larval hatching of the monogenean Diplectanum aequans, gill parasite of sea bass Dicentrarchus labrax, was studied in relation to different prophylactic treatments. Groups of eggs of D. aequans were submitted to different in vitro treatments: formalin (300 and 100 L L^–1 per 1 hour), Neguvon^® (trichlorphon 0.2 mg L^–1 per 48 hours) and dehydration for 4 and 8 hours. Percentages of hatched larvae, aborted larvae and undeveloped embryos were estimated in comparison with the control group. Results showed that 300 L L^–1 formalin and dehydration treatments were able to reduce larval hatching significantly, while Neguvon^® and 100 L L^–1 formalin treatments had no effect.     

Spawning induction and seed production of Eastern little tuna,Euthynnus affinis (Cantor, 1849), in the post‐ and pre‐spawning seasons by hormonal treatment in a semi‐closed recirculation system with elevated temperature

We previously established a method for spawning induction in Eastern little tuna (ELT) Euthynnus affinis (Cantor, 1849) by administering a gonadotropin‐releasing hormone analog (GnRHa) during the natural spawning season in Japan (August–October). In order to establish seed production of ELT in the off‐spawning season, we first conducted three spawning induction trials by GnRHa administration from October 2011 to January 2012 using ELT broodstock (2 years old; three females and four males) maintained in a 10‐m³ tank with a semi‐closed recirculation system and static elevated temperature. Average water temperature and daily egg production in three trials lasting 11–15 days were 27.0 ± 0.09°C and 268 173 eggs (Trial 1), 27.0 ± 0.11°C and 277 9098 eggs (Trial 2), and 25.5 ± 0.39°C and 291 113 eggs (Trial 3) respectively. Mean fertilization rate and mean hatching rate were 70.4% and 60.5% (Trial 1), 83.9% and 79.6% (Trial 2), and 62.5% and 57.4% (Trial 3) respectively. We also succeeded in producing ELT larvae in the pre‐spawning season (April–July), although the quantity and quality of larvae produced were inferior to those produced in other calendar months. In trials involving periodic GnRHa administration during the off‐spawning seasons, hatched larvae were obtained in the 10‐m³ tank after six of nine administrations in the 2011–2012 off‐spawning season and in 16 of 19 administrations in the 2012–2013 off‐spawning season. The findings of this study demonstrated that hormonal treatment and thermal control could be used to extend the spawning period in ELT, potentially allowing larval production in the post‐ and pre‐spawning seasons.     

Hatching time effect on the intra‐spawning larval morphology and growth in Northern pike (Esox luciusL.)

Based on the analysis of 11 morphometric variables of body (total length, body area and perimeter, myotome height and eye diameter) and yolk sac (length, height, area, perimeter and volume) of pike larvae, the aim of this study was to evaluate how larval size at hatching and growth of larvae hatched from single egg batches vary according to three hatching times: early, mid and late. Hatching time structures strongly pike larval morphometrics. Early hatched larvae have smaller body sizes at hatch, faster growth and higher yolk use efficiency than late hatched ones. Early hatched larvae seem to be premature and hatch at precocious developmental stage whereas late hatched individuals continue their growth within the egg shell and hatch at larger size but with lower reserves (yolk). A compensatory growth phase was observed for the early hatching pike larvae particularly during the first 5 days post hatch. Consequently, no significant difference in body parameters was recorded from day 10 post hatching whatever the hatching time. The higher growth accomplished by early hatched larvae may be related to a particular metabolic activity that converts more efficiently yolk into body tissues.     

Effect of incubation temperature on the embryonic development and yolk‐sac larvae of the Pacific red snapper Lutjanus peru (Nichols & Murphy, 1922)

The effect of incubation temperature on embryonic development and yolk‐sac larva of the Pacific red snapper Lutjanus peru were evaluated by testing the effect of 26, 28 and 30°C, as this is the natural thermal interval reported during the spawning season of Pacific red snapper in the Gulf of California, Mexico. Sixteen developmental stages were observed. The incubation temperature affected the rate of development and time to hatching, being shorter at 30 than at 26°C, but no significant effect (P < 0.05) on larval length at hatching was registered. The depletion rate of yolk sac and oil globule was affected by incubation temperature particularly during the first 12 h post hatching (hph). At the end of the experiment (48 hph), significantly (P < 0.05) larger larvae were recorded at 26°C (TL = 3.22 ± 0.01 mm) than at 28° (TL = 3.01 ± 0.02 mm) and 30°C (TL = 2.97 ± 0.05 mm). Incubation of newly fertilized eggs at 26°C produces larger larvae, which may help to improve feeding efficiency and survival during first feeding.     

Off-Season Spawning of Eurasian Perch Perca Fluviatilis

This study was designed to determine the effects of the selection for body weight and of two final warming durations during an out-of-season reproductive cycle on gonad recrudescence, plasma steroid levels and spawning in Eurasian perch Perca fluviatilis males and females. Two different bodyweight classes (200–250 and 350–400 g) and two different warming periods (1 or 2 months) were studied.At the end of the chilling period, a significant effect of the selection for body weight was observed on gonad development whereas fish were the same age. According to higher levels in testosterone (12 ng mL^–1), a better development was recorded for smaller breeders (P < 0.05). A short water warming period up to 14°C (1 month v.s. 2 months) allowed higher rates of females with completed gonadogenesis (33–38% v.s. 7–8%) and spawning (31–33% v.s. 0%) (P < 0.05). Spontaneous out-of-season spawning (17 ribbons) and larvae were obtained. Relative fecundity was about 100 eggs g^–1 of female body weight. Five ribbons were partly fertilized with a rate ranging from 5 to 90%. These studies have shown that delayed spawning in Eurasian perch can be obtained by a temperature manipulation, but results are not sufficient because of low stimulation rate of females, low spawning rates and high heteroge-neity of fertilization rates. The effects of other environmental factors like photoperiod should be tested in further experiment to improve the protocol inducing out-of-season spawning production.     

Is batch variability in hatching time related to size heterogeneity and cannibalism in pikeperch (Sander lucioperca)?

Larval size heterogeneity is known to induce cannibalism, and procedures to avoid larval size differences are consequently implemented already during egg incubation and hatching. We investigated the relation between larval development variability, size heterogeneity and cannibalism in pikeperch. Larvae were sorted into five groups according to the time of hatching during a 65‐h period. The larvae with different times of hatch were then reared separately or together during an 18‐day period. Late hatched larvae were longer (P=0.003) and had less yolk remaining (P<0.001) than early hatched individuals at the time of hatching. However, on 11 days post fertilization, the late‐hatching larvae tended to have larger yolk reserves than earlier hatched individuals (P=0.07). Furthermore, the next day, a lower proportion in the late fraction had switched to exogenous feeding (P=0.024). That larvae with a late hatching time developed slower suggests a positive relationship between the hatching time and the embryonic developmental rate. However, differences in the length and available yolk reserves at hatching between larval fractions with different hatching times suggest that hatching is not strictly associated with a specific developmental stage, and that factors other than the development rate of the embryo are involved in the timing of hatching.     

Embryonic and larval development of black skirt tetra (Gymnocorymbus ternetzi,Boulenger, 1895) under laboratory conditions

The embryonic and larval development of black skirt tetra, Gymnocorymbus ternetzi, are described under controlled laboratory conditions. In addition, major histomorphological changes and the allometric growth patterns during larval development have been described. The laboratory‐reared broodstock, that is 1 year of age, were spawned. Hatching occurred 20–21 h after spawning at 24 ± 0.5°C. The cleavage was finished in 2 h and the early blastula stage occurred at 2:04 hours after spawning. The gastrulation started at 3:20 hours and 30% epiboly was observed at 3:34 hours after spawning. Eight‐somite stage was observed at 08:33 hours. And embryonic developmental stage was completed at 21 h after spawning. The newly hatched larvae were 1442 ± 14.3 μm in mean total length (TL). The mouth opened at 3 days after hatching (DAH). The yolk sac had been totally absorbed and the larvae started to swim actively within 3–4 days. Notochord flexion began at 11 DAH. The metamorphosis was completed and the larvae transformed into juveniles at 32 DAH. In this paper, the full developmental sequence from egg to juvenile of G. ternetzi is described for the first time.     

Chemical treatment of lobster eggs against epibiotic bacteria

Eggs of the European lobster, Homarus gammarus (L.), were exposed to malachite green (5, 10, 15 mg 1^–1: 10 min), glutaraldehyde (50, 100, 150 mg 1^–1: 3 min) and iodine as Buffodine^TM (50, 100, 150 mg 1^–1: 10 min). The efficiency of the treatments was tested by incubating eggs individually in wells of multiwell dishes with TSB agar for 14 days after exposure. In order to find any effect on viability, batches of 30 eggs from each of three females were incubated artificially in a recirculation system for 19 days and repeatedly exposed to the disinfectants. Iodine as Buffodine (150 mg 1^–1) was the only treatment that resulted in a significant decrease of the bacterial growth on lobster eggs, but the treatment also resulted in inhibited hatching compared with the control group. Thus, our results indicate that treatment with 150mg^–1 iodine as Buffodine could be a strategy for reducing bacterial growth on lobster eggs when massive egg mortality due to bacteria is otherwise unavoidable. The treatment could, however, lead to decreased viability of larvae due to inhibited hatching.     

Utilization of lipids by Dentex dentex L. (Osteichthyes, Sparidae) larvae during lecitotrophia and subsequent starvation

Total lipids, lipid classes and their associated fatty acids were measured in developing eggs, yolksac larvae and starving larvae (from fertilized egg to day 9 after hatch) of the common dentex Dentex dentex (L., 1758). The larvae of common dentex during lecitotrophia and subsequent starvation consumed 1.6 g of total lipid per larvae per day. The overall decrease was mainly due to utilization of the major neutral lipids, TAG and SE (0.5 and 0.6 g larvae^–1 day^–1, respectively) which was 3.4-fold greater than that of the the major phosphoglycerides (primarily PC by 0.2 g larvae^–1 day^–1). There was net synthesis/conservation of PE during the first half of the study period before it decreased rapidly (0.2 g larvae^–1 day^–1) during the second half. PUFAs were principally catabolized (468.6 ng larvae^–1 day^–1), primarily 22:6(n-3), 20:5(n-3) and 20:4(n-6) (221.8, 58.5 and 12.1 ng larvae^–1 day^–1, respectively). Saturated and monounsaturated fatty acids were also utilized (227.2 and 256.7 ng larvae^–1 day^–1, respectively), principally 16:0 and 18:1(n-9) that were both consumed at 149.8 and 156.7 ng larvae^–1 day^–1, respectively. The rank order of utilization of fatty acids (ng larvae^–1 day^–1) by D. dentex larvae from total lipids, PC and TAG coincided with the order of abundance of the different fatty acids in the respective lipid fractions. However, in PE, the most abundant fatty acid, DHA, was relatively conserved and 16:0, the second most abundant fatty acid, was catabolized to the greatest extent. D. dentex showed a pattern of lipid metabolism during early development similar to that of marine larval fish from temperate waters whose eggs contain high levels of total lipids, including an oil globule, and which preferentially utilize neutral lipids as the primary energy source.     

Egg and larval quality,and egg fatty acid composition of Eurasian perch breeders (Perca fluviatilis) fed different dietary DHA/EPA/AA ratios

In Eurasian perch (Perca fluviatilis), the variability in spawning quality is a major limiting factor for successful production, especially when breeders are fed with an artificial diet. The influence of the dietary DHA/EPA/AA ratio on the egg and larval quality and on the fatty acid and lipid class composition of eggs has been investigated in perch broodstock. Two experimental diets (16% lipids) with two different DHA/EPA/AA ratios, D1 (3/2/2) and D2 (23/9/1), were compared with a natural diet consisting of cultured carp juveniles, CC (10/10/1) and with a commercial diet for salmonids, CDS (14/16/1). Percentages of fertilization and hatching were comparable between fish fed D1, D2 and CC, with the highest hatching rate observed for D1 (63.5 ± 3.8%). These diets supported better values than the CDS. Larval survival and TL50 observed after osmotic stress were higher for the D1 group, followed by larvae produced by fish fed D2 and CC. Larvae from fish fed D1, D2 and CC were significantly more robust than larvae from the CDS group. Differences were observed regarding the fatty acid (FA) profile in the eggs, which was related to the dietary FA composition. The results indicate that a ratio of 3/2/2 seemed to be effective for obtaining eggs and larvae of good quality.     

Incorporation of yolk fatty acids into body lipids of goldfish (Carassius auratus L.) larvae raised at two different temperatures

In five separate experiments, eggs from a single female goldfish were fertilized at 20°C. They were incubated at 22°C for 6 hours, after which some of the eggs were transferred to 13°C. When a defined post-hatch developmental stage was reached, lipid extracts were prepared from larvae, both with yolk sacs intact and after removal of the yolk sac by dissection. Other larvae were sampled at yolk exhaustion. Gas chromatographic analysis of fatty acid profiles revealed that larvae incorporated 16:0, 18:0, 20:4 (n–6) and 22:6 (n–3) into their tissues in proportions higher than those present in the eggs from which they were derived. At 22°C, these trends were particularly apparent at yolk exhaustion. At 13°C, proportions of polyunsaturated fatty acids in the bodies of newly hatched larvae were higher than those in the 22°C larval bodies. Monounsaturated fatty acids were preferentially depleted during development, especially in larvae from high quality eggs. No dependence of egg quality, as assessed by larval viability at 22°C, on total egg lipid mass or fatty acid composition was found. Larvae from the lowest quality eggs showed a reduced preference for incorporation of (n–3) polyunsaturated fatty acids into their tissues.     

Induction of Out-of-Season Spawning of Pikeperch, Sander Lucioperca (L.)

Pikeperch were induced to spawn 3 months prior to the natural spawning period through photothermal and hormonal stimulation. Females (five specimens in each group) were stimulated with injection of human chorionic gonadotropin (hCG) once (200 IU kg^–1), twice (200 IU kg^–1, second dose after 48 h–400 IU kg^–1) or three times (200 IU kg^–1, after 24 h–200 IU kg^–1 and after another 24 h–200 IU kg^–1). The control group was injected once with 0.9% NaCl. The males were stimulated with a single hormone dose of 200 IU kg^–1. Eggs were obtained from all the hormonally treated fish. None of the control group females, which were only stimulated photothermally, ovulated any eggs. The time of ovulation was 66–71 h following the first injection, and the eggs viability until the eyed stage (from 71.5 to 77.5%) did not depend on the number of hormone doses (P > 0.05). The out-of-season spawning method described in this paper could be used to provide pikeperch larvae for intensive culture systems (recirculating water systems) before natural spawning season and to produce larger-sized pikeperch fingerlings for stocking.     

Reproductive behaviour and early development in yellowtail kingfish (Seriola lalandi Valenciennes 1833)

The spawning behaviour of wild caught brood stock as well as early egg and larval development were studied in yellowtail kingfish (Seriola lalandi). Spawning occurred naturally in the austral spring/summer (November-February) when the seawater temperature was above 17 °C. Courtship behaviour involved one male and female, and consisted of a high-speed pursuit punctuated by stalling, nipping and touching. This lasted for approximately 0.5-1.5 h until, immediately prior to spawning, the male would nip at the female gonoduct, presumably to induce spawning. At this stage, in 50% of spawns, a second male would become involved. The release of gametes involved frenzied circling behaviour near the bottom of the tank and lasted approximately 22 s. Spawning occurred in the early daylight hours at the start of the spawning season, but shifted to around dusk in the latter part. Spawned eggs were positively buoyant, had a high fertilisation rate (> 99%), ranged 1.33-1.50 mm in diameter with a single oil droplet 0.30-0.33 mm diameter, and developed in a similar manner to that described in congenerics. Egg viability within the floating fraction was visually determined to be 74% ± 17% over the entire reproductive season. Indistinct cell margins and asymmetrical cleavage were the most common blastomere deformities observed. Egg and oil droplet volume were found to decrease by 15-20% over the spawning season, though no relationship was found between visually assessed egg viability and date. Egg incubation trials between 16 and 24 °C indicated that temperature accelerated the time to hatch by a Q₁₀ of 5.0. While larvae were found to hatch at a smaller length with a larger yolk sac and oil droplet at warmer incubation temperatures, there was little difference in the maximum larval length reached at the onset of first feeding among the rearing temperatures used. It is proposed that the reason for this was that higher incubation temperatures accelerated the hatching process faster than the rate of tissue deposition. The findings from this study are discussed in terms of the biological significance and implications for the larviculture of this species.     

Biological Performance of Red Porgy (Pagrus pagrus) Larvae under Intensive Rearing Conditions with the use of an Automated Feeding System

Pagrus pagrus is one of the promising species for the industry in the Mediterranean but its rearing is still far from satisfactory. In the present work, the conditions and results of larval rearing with the use of an automatic feeding system are presented. Eight populations were reared for 20 days with the pseudo green water methodology in two successive trials. Larvae were fed enriched rotifers during the entire rearing period, a mixed diet of rotifers and Artemia (Instar I when larvae reached 5 mm in length, replaced by Instar II after larvae reached 5.5 mm) and a formulated diet (after day 16 post hatching). A computerized system for feeding management was used. A total of 388,000 eggs were incubated and after 20 days, 237,973 larvae (TL = 6.96 ± 0.17 mm) were produced with a survival rate of 61.4 ± 6.3%. Individuals grew in terms of wet weight with an exponential rate of 0.167 ± 0.008 daily (R ²= 0.983) with no variations between replicates. Mean individual daily consumption at first feeding (day 4-post hatching), was 0.20 ± 0.06 mg of food (0.03 ± 0.01 dry weight), that is, 180–300 rotifers, while on day 20, consumption increased by 10 (2.081 ± 0.106 mg and 0.276 ± 0.014 in wet and dry weight, respectively). When compared with sea bream, consumption was higher by approximately 3–4-fold. The mean food conversion ratio of the experimental period was 4. In comparison with sea bream larviculture, food consumption of red porgy is higher, and a different feeding strategy is required to satisfy the requirements of the larvae.     

Drinking activity of the newly hatched larvae of codGadus morhua L.

The drinking rate of cod larvae 1–7 days post hatching was measured from the uptake of³H-labelled dextran (MW = 70000) admixed in the incubation seawater (34 ppt 5°C). The drinking rate increased gradually from 0.15% to 0.59% of the larval body weight on day 1 and day 7 respectively. This increase in drinking rate correlated with an observed decrease in the volume of the yolk sac and its water store. Autoradiographs showed the labelled dextran to be confined to the intestine. Electron micrographs showed an open mouth communicating with the oesophagus and the intestine in cod larvae at the time of hatching. Chloride cells were present on the opercular folds but not on the vestigial, developing gills. The data indicate that the water acquisition mechanism of larval cod is similar to that of adult marine fish.     

The Scale‐up of Spotted Rose Snapper,Lutjanus guttatus,Larval Rearing at Mazatlan,Mexico

The scale‐up of spotted rose snapper, Lutjanus guttatus, larval rearing is described. Fertilized eggs (480,000) were obtained from a 1‐d harvest of a natural spawning captive broodstock acclimatized for 1 yr and 6 mo in two fiberglass tanks (18 m³). Fourteen hours after spawning, 89.6% of the collected eggs were floating, of which 96.2% were transparent with live embryos. Incubation at 25–26 C lasted 21 h, with 90.2 ± 2.1% hatching percentage of normal larvae. The percentage of viable larvae at 48 h after hatching was 79.7 ± 1.9%. Initial stocking density was 10.4 ± 1.0 larvae/L 2 days after hatching (d.p.h.). A total of 22,600 juveniles (1256 ± 170 juveniles/m³) were harvested from six 3‐m³ cylindrical fiberglass tanks. Average survival was 12.1 ± 1.1%. Final mean length and weight were 5.5 ± 0.05 cm and 2.24 ± 0.04 g, respectively. Growth expressed in total length was TL = 2.1476e^0.0543t (R² = 0.9911). Final mean biomass and condition factor were 2.8 kg/m³, 12.3% and 1.346. General length‐weight ratio was W = 0.05460 LT^2.2306.     

Development of osmoregulatory tissues in the Lake van fish (<Emphasis Type="Italic">Alburnus tarichi</Emphasis>) during larval development

Lake Van is one of the largest alkaline lakes worldwide and Lake Van Fish (Alburnus tarichi Güldenstädt, 1814) is the only vertebrate species inhabiting it. Lake Van Fish is an anadromous species that migrates to the streams (salinity 0.02%, pH 8.42) flowing into Lake Van (salinity 0.22%, pH 9.8) during the spawning period (April–July). Following spawning, fish return to Lake Van while larvae remain in fresh water. This study examined the development of osmoregulatory organs and the distribution of ionocytes in Lake Van Fish larvae adapting to the highly alkaline water characterizing the lake. Ionocytes were marked immunohistochemically and observed in whole mounts with immunofluorescence staining using the Na⁺/K⁺ ATPase antibody. Ionocytes were first identified in the yolk sac membrane and skin, and then in the gills, digestive tract, and kidneys of larvae. The number of ionocytes on yolk sac membrane and skin decreased during larval development, indicating ionocytes on these tissues have a role in larvae osmoregulation. Larvae hatched from eggs in stream waters die when transferred to Lake Van water but survived in lake water diluted with deionized water. Thus, larvae need to go through certain alterations at the cellular and organ levels in order to adapt to the conditions of Lake Van water, indicating they do not enter this lake immediately after hatching.