Effects of temperature and photoperiod on ovarian recrudescence in the cyprinid fish Mirogrex terrae-sanctae

Mirogrex terrae-sanctae is a commercial, sardine-like cyprinid fish in the Sea of Galilee. The fish spawns from December to the end of March. Females collected at the quiescent phase of the cycle (June) were exposed to four combinations of photoperiod and temperature for 3 months. The highest GSI (2.44 ± 0.22) was found in females exposed to simulated winter conditions (16°C, 9L) while the lowest GSI (0.65 ± 0.04) was found in fish exposed to simulated summer conditions (27°C, 14L). The GSI of fish maintained at the unnatural combination of 16°C, 14L (1.30 ± 0.09) did not differ from that of fish maintained at the other unnatural combination of 27°C, 9L (1.07 ± 0.11). However, histological examinations revealed that vitellogenic oocytes were present only in fish maintained at low temperature regimes, which implies that the growth of the ovarian mass in the 27°C, 9L group was due mainly to oogonial proliferation and that vitellogenesis was inhibited in Mirogrex at a high temperature. Both protein and calcium concentrations increased in the serum of fish following injections of oestradiol-17β. The increase of calcium was evident only in the non-filtrable (bound) fraction, suggesting that the total serum calcium above 13 mg% could be used as an indirect parameter for circulating vitelloprotein. Fish collected at the quiescent phase were injected with oestradiol (50 μg/fish c. 15 g body weight) and subsequently transferred to tanks maintained at either 12°, 20° or 27°C. The total serum calcium increased to 23.0 ± 1.75 mg% in the fish maintained for 8 days at 20°C. This was significantly higher than the concentration of calcium in the serum of fish maintained at either 12°C (7.8 ± 0.52) or 27°C(l1.8 ± 0.77). This implies that the temperature may affect ovarian recrudescence by modulating the response of the liver to oestrogen, allowing the oestrogen-induced vitellogenesis to be relatively high in the cool environment during winter.     

The histology of mature gonads of Oreochromis (Nyasalapia) karongae (Trewavas)

Histology of gonads of Oreochromis karongae was undertaken to study internal cell characteristics during maturation. This study was necessitated by low spawning output of the fish species. Several oocyte stages, ranging from primary forms to vitellogenesis, suggest that the maturation was generally succesfully attained in the fish ponds. Pre‐vitellogenesis oocytes (oogonia to perinuclear stage) and more advanced vitellogenesis (primary vesicle to tertiary yolk vesicle) oocyte stages were all found in the same gonads. However, there were some discontinuities observed during stages 3 and 4, suggesting selective maturation. Failure of gonads to mature normally is attributed to an ecological crunch that was in a previous study associated with environmental factors. Atretic oocytes were also recorded in the same gonads, a sign that some oocytes failed to mature normally. This indicates insufficient stimuli for normal gonad development. Several stages of spermatogenesis (spermatocytes, spermatid and spermatozoa) were also found in the same gonads. Selective recrudescence was more pronounced in O. karongae because generally less oocytes attained final maturation stages compared to Oreochromis niloticus and other tilapias. This could be the main reason for low natural breeding that has been observed in both wild and captive stocks, and led to the abandonment of its use in aquaculture. This study corroborates findings of previous studies that depended solely on external gonad characteristics. Histology provides conclusive evidence from internal cell characteristics that other techniques are unable to show.     

Determination of batch fecundity in Uranoscopus scaber Linnaeus, 1758 from the Aegean Sea, Turkey

This work focuses on reproductive biology aspects of Uranoscopus scaber from the coast of the Aegean Sea. In total, 175 U. scaber specimens were collected in Izmir Bay using trawl nets from January to December 2005. The reproduction period according to gonadosomatic index (GSI) values extents from March to September, with a peak in May (6.32 ± 1.2). Oocyte diameters taken from 59 mature gonads ranged from 62 to 1860 μm. Maximum batch fecundity (BF) was 58091 (mean: 11556 ± 1445) in large oocytes in May, and 8408 (mean: 2821 ± 945) in hydrated oocytes in September. The BF increased with total length and body weight, excluding the ovaries. Total length (TL)–BF relationships and fish weight–BF relationships were BF = 0.0034TL^4.6248 (r = 0.47) and BF = 59.166W?3101 (r = 0.49), respectively, according to the oocyte size frequency method. There was a linear relationship between BF and fish size.     

Cyclic variations of gonad development of snow trout,Schizopyge niger from the river Jhelum of Kashmir Himalaya

The aim of the present study was to know the cyclic changes in gonadal maturation and to investigate the various developmental stages of oocytes and testicular germ cells of snow trout, Schizopyge niger. A total of 316 fish specimens were procured from three sites of river Jhelum during November 2016 to October 2017 and the histological analysis of their gonad was carried out. The results of the present study indicates that the male population was found significantly (p < 0.05) higher compared to female population and were reported as (1.6:1; M:F). The highest mean GSI value was recorded in male (12.24 ± 0.89) during the month of March and for female (13.74 ± 0.92) also in the month of March, which indicated that the gonadal development reached its peak during this month. The highest mean oocyte diameter was 1.04 ± 0.20 mm observed in March, showing that the oocyte reached its maturity during this month. Histological study of ovary revealed the evidence of vitellogenic yolk granule stage and ripe mature stage from December to April. While in case of male, four stages of spermatogenesis were distinguished and spermatozoa were highly abundant in the months of March and April. Mean ± SE of absolute fecundity was 3,224 ± 421 eggs and also, the relative fecundity was 69 egg/g body weight. Based on the results of GSI and the analysis of gonad histology, it was observed that the peak breeding season of S. niger occurred in the month of March. It was also noted that spawning period of this fish begins in the month of February and lasts till April with a peak breeding season in the month of March. The present study will provide a baseline information about the appropriate peak breeding of the fish, which will be useful for artificial breeding as well for prevention of fishing from natural habitat.     

Gonadal maturation,fecundity, spawning,and timing of reproduction in the mud snail,Cerithidea cingulata,a pest in milkfish ponds in the Philippines

Summary

Gonadal maturation, spawning, fecundity and timing of reproduction of the snail Cerithidea cingulata in a brackish water pond in Molo, Iloilo, Philippines, are described. Snails 4–41 mm in shell length were sampled monthly from May 1997 to May 1998; 25% were <25 mm, 67% were 20–30 mm, and 8% were >30 mm. The sexes are separate and could first be distinguished at 15 mm. Males are aphallic, have narrower shells than females of the same length, and have bright yellow-orange testes overlying the digestive gland deep inside the shell. Females have more robust shells, an ovipositor at the right side of the foot, and yellow-green ovaries overlying the digestive gland. The sex ratio was one male to two females in the pond population studied. Gonadal maturation was monitored by means of gonadosomatic index (GSI, gonad weight as a percent of visceral weight); maturation stages were based on the gonad appearance (immature, developing, mature) and histology (immature, developing, mature, redeveloping). GSI increased with snail size, and reached 16% in a 33-mm female. The smallest mature males and females were 18–19 mm, and most snails >20 mm were mature, spawning, or redeveloping. Histological sections showed all stages of gametogenesis in mature male snails. The oocyte size-frequency distributions in mature females showed mostly mature oocytes and secondary oocytes, but also oogonia and primary oocytes. GSI and the frequency of snails at different maturation stages varied over the year. Both GSI and the frequency of mature snails were highest during the summer months, April to August. Nevertheless, mature snails occurred throughout the whole year, as did mating and egg-laying. Fecundity (= number of oocytes >70 pμ) increased with size in mature females 2041 mm; an average 25-mm female produced about 1,500 oocytes and larger females produced a maximum of about 2,500 oocytes. Eggs strings laid on the pond bottom were 45–75 mm long; an average 64-mm string contained 2,000 eggs 210+20 pm in diameter. The density of eggs strings was highest (80–120/m²) during March-September. Eggs hatched after 6–7 d into planktonic veligers, which in turn settle on the pond bottom 11–12 d later as juveniles. Juveniles 2–6-mm long were most abundant in the pond during August-October.     

Reproductive biology ofAwaous guamensis, an amphidromous Hawaiian goby

Synopsis Spawning season, size at first reproduction, oocyte maturation, and fecundity ofAwaous guamensis, an amphidromous Hawaiian goby, were studied from June 1989 through May 1991 in the Wainiha River, Kau'ai, Hawai'i. Female fish larger than 73 mm standard length (SL) had mature gonads from August through December in 1989 and 1990. Gonadosomatic index (GSI) values for mature females ranged from 0.2 to 14.5 during the spawning season. Male fish larger than 64 mm SL had elevated GSI values from June 1989 through December 1989 and from August 1990 through December 1990. Mature sperm were found in two male fish collected in January and February. GSI values for mature males ranged from less than 0.01 to 4.0 in the spawning season. Size-frequency distributions of measurements of vitellogenic oocyte diameters and microscopic observations of oocytes indicated this species has group-synchronous oocyte development. Ovarian maturation stages examined over a 29-month period suggest that members of the stock spawned at different times within the spawning season, although mass spawning events have been documented for this species. Estimates of clutch sizes from nests measured in situ were comparable to estimates of potential fecundity from in vitro examination of ovaries, and indicated that female fish deposited an entire clutch during a spawning event. No evidence for multiple spawning by an individual fish in a single season was found. However, microscopic observations of brown bodies in some ovaries suggested that individual fish probably spawn more than once in a lifetime.     

Effect of developmental temperature on swimming performance of zebrafish (<Emphasis Type="BoldItalic">Danio rerio</Emphasis>) juveniles

It is widely known that water temperature affects the swimming capacity of fish. But the effect of the rearing temperature on the swimming ability of the fish at later stages, has not had similar attention. In this study, four populations of zebrafish, were reared in different water temperatures (22, 25, 28 and 31°C) and after being acclimatized in a common temperature (26.5°C) for over a month, they were subjected to swimming trials in order to evaluate the maximum relative critical velocity (RU _crit ) in each case. Fish that were reared in 22°C showed statistically significant lower performance than the ones reared in 31°C (7.72 ± 0.17 vs. 8.79 ± 0.28, means ± S.E.). Possible explanations for the observed differentiation could be the effect of early life temperature on fish muscle ontogeny or on body shape.     

Reproductive cycle of gilt sardine,Sardinella aurita,Valenciennes 1847, in the eastern middle Adriatic Sea

The annual reproduction cycle of gilt sardine, Sardinella aurita, based on gonad maturity stages, gonad weight and gonadosomatic index was the subject of this study. A total of 2033 gilt sardines (983 males, 1021 females and 29 undetermined) were analysed. Fish were collected monthly from commercial purse seiners between November 2007 and January 2009 in the eastern middle Adriatic Sea (mesh size 8 mm/bar length/; sampling: five boats per month). Based on the monthly evolution of gonad maturation stages, gonad weights and gonadosomatic index, the peak spawning season was determined to be from June to August. Variations in sea surface temperature (SST) coincided with monthly variations of the gonadosomatic index. Highest monthly average values for both analysed parameters were recorded in July (GSI = 3.38; T = 26.5°C). Fifty per cent (L₅₀) of males and females reached sexual maturity at TL 15.8 cm and at 16.6 cm, respectively. Absolute fecundity ranged from 8458 to 48 032 (mean 34 565 ± 10 310), whereas relative fecundity was from 171 to 722 (mean 385 ± 104.35). Mean value of the oocyte size was 0.53 ± 0.10 mm.     

The critical thermal limits for the bullhead, Cottus gobio, from three populations in north-west England

1. The objective was to determine the thermal limits for feeding and survival in the bullhead, Cottus gobio, using juveniles (total length 20–30 mm, live weight 0.5–1.5 g) from one population and adults (50–70 mm, 3.5–5.5 g) from three populations. 2. Fish were acclimated to constant temperatures (3, 7, 10, 15, 20, 25 or 27 °C) and the temperature was then changed at a rate of 1 °C /30 min to determine the critical limits for feeding, survival over 7 days (incipient lethal temperature), or survival for 10 min or less (ultimate lethal temperature). The rate of 1 °C/30 min was the optimum value from preliminary experiments, using nine rates from 0.5 °C/48 h to 18 °C h^?1. As values for adults were not significantly different between populations, they were pooled to provide arithmetic means (with 95% CL) for the thermal limits at each acclimation temperature. 3. Feeding limits increased with acclimation temperature to upper and lower mean values (± 95% CL) of 26.5 ± 0.16 °C and 4.2 ± 0.20 °C for adults, 26.6 ± 0.59 °C and 5.0 ± 0.55 °C for juveniles. Incipient lethal levels defined a tolerance zone within which fish survive indefinitely; upper limits increased with acclimation temperature to a plateau of 27.6 ± 0.22 °C for adults and 27.5 ± 0.47 °C for juveniles, lower limits increased from near 0 °C to 2.5 ± 0.31 °C for adults and 2.7 ± 0.47 °C for juveniles. Ultimate lethal levels increased with acclimation temperature to a plateau of 32.5 ± 0.24 °C for adults and 32.6 ± 0.46 °C for juveniles, whilst the lower limits increased from near 0 to 0.9 ± 0.29 °C. Upper feeding, incipient and ultimate lethal values were significantly lower for juveniles than those for adults at acclimation temperatures < 20, < 20 and < 15 °C, respectively. 4. The thermal tolerance of bullheads was slightly lower than that of stone loach, similar to that of juvenile Atlantic salmon and higher than that of brown trout; the thermal limits for feeding were much wider than those for salmon or trout.     

Comparative study of reproductive biology in single- and multiple-spawner cyprinid fish. I. Morphological and histological features

To clarify the dynamics and regulation of oogenesis in single- and multiple-spawning cyprinid fish with group-synchronous oocyte development, a multidisciplinary approach to their reproduction was undertaken using three species from the River Meuse (Belgium): the roach Rutilus rutilus as a single spawner, and the bleak Alburnus alburnus and the white bream Blicca bjoerkna as multiple spawners. The gonadosomatic index (GSI) and histomorphometric changes (distribution of oocyte size, relative proportion of the various oocyte stages) in the ovary are compared. Different patterns of GSI and oocyte growth were observed both between the single- and multiple-spawner fish and between the two multiple spawners. Maximum GSIs were higher in roach (21%) than in bleak and white bream (17.7 and 14.5%, respectively), and compared to the rapid decline of GSI in the roach population, the GSI of multiple spawners decreased progressively during the spawning season. In roach, a short gonadal quiescent period and an early onset of vitellogenesis was recorded from late summer onwards whereas, in bleak and white bream, exogenous vitellogenesis was not systematically observed before winter. A protracted spawning season and/or a low water temperature in autumn are hypothesized to explain this long period of gonadal quiescence. In bleak, during the spawning season, the oocytes recruited arose from the stock of endogenous vitellogenesis and attained the final maturation stage very rapidly. This recruitment occurred during the whole spawning season. In white bream, the differentiation of vitellogenic oocytes from smaller oocytes was completed before the onset of the spawning season. During the spawning period, the proportion of vitellogenic oocytes decreased progressively whereas the percentage of oocytes in the final maturation stage remained approximately constant.     

Thermotolerance of IVM‐derived bovine oocytes and embryos after short‐term heat shock

A series of experiments were designed to study the effect of elevated temperatures on developmental competence of bovine oocytes and embryos produced in vitro. In experiment 1, the effect of heat shock (HS) by a mild elevated temperature (40.5°C) for 0, 30, or 60 min on the viability of in vitro matured (IVM) oocytes was tested following in vitro fertilization (IVF) and culture. No significant difference was observed between the control (39°C) and the heat‐treated groups in cleavage, blastocyst formation, or hatching (P > 0.05). In experiment 2, when the HS temperature was increased to 41.5°C, neither the cleavage rate nor blastocyst development was affected by treatment. However, the rate of blastocyst hatching appeared lower in the HS groups (13% in control group vs. 3.9% and 5.6% in 30 min and 60 min, respectively; P < 0.05). When IVM oocytes were treated at 43°C prior to IVF (experiment 3), no difference was detected in blastocyst and expanded blastocyst development following heat treatment for 0, 15, or 30 min, but heat treatment of oocytes for 45 or 60 min significantly reduced blastocyst and expanded blastocyst formation (P < 0.05). In experiment 4, the thermotolerance of day 3 and day 4 bovine IVF embryos were compared. When embryos were pre‐treated with a mild elevated temperature (40.5°C) for 1 hr, and then with a higher temperature (43°C) for 1 hr, no improvement in thermotolerance of the embryos was observed as compared to those treated at 43°C alone. However, a higher thermotolerance was observed in day 4 than day 3 embryos. In conclusion, treatment at 43°C, but not 40.5°C or 41.5°C significantly reduced oocyte developmental competence. An increase in thermotolerance was observed from day 3 to day 4 of in vitro embryonic development, which corresponds to the maternal to zygotic transition of gene expression in bovine embryos. Mol. Reprod. Dev. 53:336–340, 1999. © 1999 Wiley‐Liss, Inc.     

Studies on chilling sensitivity of zebrafish (Danio rerio) oocytes

Human activity in the last few decades has had a devastating effect on the diversity of fresh water and marine fish. Further decline of fish population may have serious economic and ecological consequences. One of the most promising techniques to preserve fish population is to cryopreserve their germ cells. Cryopreservation has been successfully applied to fish sperm of many species, but there has been no success with fish embryo cryopreservation and fish oocyte cryopreservation has never been studied systematically. The aim of this study is to investigate the chilling sensitivity of fish oocytes. Experiments were conducted with zebrafish stage III (vitellogenic) and stage V (mature) oocytes, which were chilled at 10, 5, 0, -5 or -10 degrees C for 15 or 60 min using a low temperature bath. Control oocytes were kept at room temperature at 22 degrees C. Oocyte viability was assessed using three different methods: trypan blue staining (TB), thiazolyl blue tetrazolium bromide (MTT) staining and observation of germinal vesicle breakdown (GVBD). The results showed that zebrafish oocyte are very sensitive to chilling and their survival decreased with decreasing temperature and increasing exposure time periods. Normalised survivals assessed with TB staining after exposure to 0, -5 or -10 degrees C for 15 or 60 min were 90.1+/-6.0, 77.8+/-7.6, and 71.2+/-9.3%, and 60.2+/-3.8, 49.6+/-6.7, and 30.4+/-3.0%, respectively. The study found that the sensitivity of viability assessment methods increase in the order of MTT < TB < GVBD. It was found that stage III oocytes were more susceptible to chilling than stage V oocytes, and that individual female had a significant influence (p < 0.0001) on oocyte chilling sensitivity. Zebrafish oocyte chilling sensitivity may also be one of the limiting factors for development of protocol of their cryopreservation.     

Improvement of gamete quality and its short-term storage: an approach for biotechnology in laboratory fish

In fish, in vitro fertilization is an important reproductive tool used as first step for application of others biotechniques as chromosome and embryo manipulation. In this study, we aimed to optimize gamete quality and their short-term storage from the yellowtail tetra Astyanax altiparanae, for future application in laboratory studies. Working with sperm, we evaluated the effects of spawning inducers (carp pituitary gland and Ovopel® [(D-Ala6, Pro9-NEt) – mGnRH+metoclopramide]) and the presence of female on sperm motility. Additionally, we developed new procedures for short-term storage of sperm and oocytes. Briefly, sperm motility was higher when male fish were treated with carp pituitary gland (73.1±4.0%) or Ovopel® (79.5±5.5%) when compared with the control group treated with 0.9% NaCl (55.6±27.2%; P=0.1598). Maintenance of male fish with an ovulating female fish also improved sperm motility (74.4±7.4%) when compared with untreated male fish (42.1±26.1%; P=0.0018). Storage of sperm was optimized in modified Ringer solution, in which the sperm was kept motile for 18 days at 2.5°C. The addition of antibiotics or oxygen decreased sperm motility, but partial change of supernatant and the combination of those conditions improve storage ability of sperm. Fertilization ability of oocytes decreased significantly after storage for 30, 60 90 and 120 min at 5, 10, 15 and 20°C when compared with fresh oocytes (P=0.0471), but considering only the stored samples, the optimum temperature was 15°C. Those data describe new approaches to improve semen quality and gametes short-term storage in yellowtail tetra A. altiparanae and open new possibilities in vitro fertilization.     

Capacitation in vitro of bovine spermatozoa by oviduct epithelial cell monolayer conditioned medium

The effect of the active capacitating factor secreted from oviduct epithelial cell monolayers (OECM) in different environments on in vitro fertilization was evaluated. Capacitation was determined as the ability of sperm to fertilize bovine oocytes in vitro. When the mTALP was supplemented with glucose during conditioning, the sperm penetration rate was significantly reduced (P ≤ 0.05) compared to the control (7% ± 1 vs. 30% ± 4). The percentages of sperm penetrated oocytes were higher following insemination in the OECM-conditioned medium derived from the early stage (48% ± 7) of the estrous cycle than in the OECM-conditioned medium derived from either mid (35% ± 2) or late stages (28% ± 3) of the estrous cycle. When the medium was supplemented with 0.1 or 0.5 μg/ml estradiol-17β during medium conditioning, sperm penetration rates increased (P ≤ 0.05) compared to the control group (55% ± 4 vs. 40% ± 3 and 54% ± 2 vs. 41% ± 3, respectively). In addition, the percentages of penetrated oocytes significantly decreased (P ≤ 0.05) following insemination when the OECM-conditioned medium was added to 0.01%, 0.05%, and 0.1% ethanol compared to the control (25% ± 4, 19% ± 2, 18% ± 3, and 45% ± 3, respectively). Sperm penetration rates significantly (P ≤ 0.01) decreased when the OECM-conditioned medium was heated to 100°C for 5 min (10% ± 1 vs. 40% ± 3). These results suggest that the active capacitating factor was sereted by the OECM and that this capacitating factor in the OECM-conditioned medium was inhibited by the presence of glucose. This factor was found to be heat-sensitive and its action was affected by ethanol. The OECM derived from the three phases of the estrous cycle as well as the presence of estradiol-17β influenced the capacity of the OECM to secrete this capacitating factor in Vitro. © 1995 wiley-Liss, Inc.     

γ-glutamyl transpeptidase of spermatozoa may decrease oocyte glutathione content at fertilization in pigs

The presence of γ-glutamyl transpeptidase (GGT) in boar spermatozoa and the potential role of the GGT at sperm penetration were examined using in vitro matured porcine oocytes. In the first experiment, GGT of boar spermatozoa was examined using a histochemical stain. GGT was detected in the midpiece and the acrosome regions of boar spermatozoa. In the second experiment, porcine oocytes matured in vitro were injected with approximately 40 pl of 10 mM HEPES solution alone or HEPES containing 0.5 U/ml GGT or 1 mM guanosine-5′-0-(3′-thiotriphosphate) (GTP-γ-S; G-protein activator). When GGT was injected into oocytes, the incidence of oocytes activated (23.7 ± 1.4%) was not different (P > 0.05) from HEPES-injected controls (24.9 ± 1.3%) at 6 h after injection. Injected GTP-γ-S, however, activated 76.0 ± 5.3% of oocytes at 6 h after injection, but extrusion of the second polar body was very low (2.8 ± 4.8%). Total content of glutathione (GSH) and glutathione disulfide (GSSG) did not differ (P > 0.05) between GTP-γ-S injected oocytes (4.2 ± 0.7 pmol/oocyte) and noninjected oocytes (4.0 ± 0.1 pmol/oocyte) at 6 h after injection. However, the total content of GSH and GSSG was lower (P < 0.01) in GGT-injected oocytes (2.1 ± 0.2 pmol/oocyte) than HEPES-injected oocytes (3.4 ± 0.2 pmol/oocyte) at 6 h after injection. In the third experiment, in vitro matured porcine oocytes were injected with about 40 pl of 10 mM HEPES solution alone or HEPES containing 0.5 U/ml GGT and then inseminated. At 12 h after insemination, the incidence of male pronuclear formation was significantly lower in oocytes injected with GGT as compared with injected control oocytes. These results demonstrated that (1) GGT was present on the surface of spermatozoa, (2) total oocyte content of GSH and GSSG was decreased by microinjection of GGT but not by that of GTP-γ-S, and (3) male pronuclear formation was inhibited in GGT-injected oocytes. These results suggest that sperm GGT may be a limiting factor for male pronuclear formation in polyspermic oocytes. © 1996 Wiley-Liss, Inc.     

Tolerance and resistance to thermal stress in juvenile Atlantic salmon, Salmo salar

SUMMARY. 1. The chief objective was to construct a thermal tolerance polygon for juvenile Atlantic salmon, Salmo salar L., using fish from four groups and two populations: two age groups from one population (0+, 1+ parr from River Leven), two size groups from the other population (slow and Fast growing 1+ parr from River Lune). 2. Fish were acclimated to constant temperatures of 5, 10, 15, 20, 25 and 27°C; then the temperature was raised or lowered at 1°C h^?1 to determine the upper and lower limits for feeding and survival over 10 min, 100 min, 1000 min and 7 days. As they were not significantly different between the four groups of fish, values at each acclimation temperature were pooled to provide arithmetic means (with SE) for the thermal tolerance polygon. 3. Incipient lethal levels (survival over 7 days) defined a tolerance zone within which salmon lived for a considerable time; upper mean incipient values increased with increasing acclimation temperature to reach a maximum of 27.8±0.2°C, lower mean incipient values were below 0°C and were therefore undetermined at acclimation temperatures <20°C but increased at higher acclimation temperatures to 2.2±0.4°C. Resistance to thermal stress outside the tolerance zone was a function of time; the ultimate lethal level (survival for 10 min) increased with acclimation temperature to a maximum of 33°C whilst the minimum value remained close to 0°C. Temperature limits for feeding increased slightly with acclimation temperature to upper and lower mean values of 22.5±0.3°C and 7.0±0.3°C. 4. In spite of different methodologies, values in the present investigation are similar to those obtained in previous, less comprehensive studies in the laboratory. They also agree with field observations on the temperature limits for feeding and survival. Thermal tolerance polygons are now available for eight species of salmonids and show that the highest temperature limits for feeding and survival are those recorded for juvenile Atlantic salmon.     

BH4 peptide derived from Bcl‐xL and Bax‐inhibitor peptide suppresses apoptotic mitochondrial changes in heat stressed bovine oocytes

Mitochondria play an important role in the integration and transmission of cell death signals mediated by the Bcl‐2 family proteins. Experiments were conducted to determine whether the anti‐apoptotic peptides BH4 domain of Bcl‐xL (TAT‐BH4) and Bax inhibitor peptide (BIP) suppresses heat stress (HS) injury in oocytes by reduction of apoptotic‐like events. Cumulus–oocyte complexes (COCs) were matured at 39°C (control) or 41°C (HS) for 21 hr then placed in maturation medium containing 0 or 100 µM BIP in water and 0 or 1 µM TAT‐BH4 in dimethyl sulfoxide (DMSO), or a combination of both peptides (BIP + BH4). Peptide effects on embryo development, DNA fragmentation, mitochondrial membrane potential (ΔΨm), and mitochondrial DNA (mtDNA) copy number were measured. All groups were fertilized and cultured in vitro at 39°C for 8 days. Compared to control, HS‐treated oocytes induced a decrease in embryo development (P < 0.05), increase in proportion of TUNEL‐positive chromatin in oocytes and blastocysts (P < 0.05), and loss of oocyte ΔΨm (P < 0.001). In the presence of BIP or BIP + BH4, development of HS‐treated oocytes into blastocysts was increased (P < 0.05). Conversely, COCs matured with TAT‐BH4 at 41°C showed reduced embryonic development (P < 0.05). Exposure of HS‐treated to each or both peptides resulted in a reduction of TUNEL frequency in oocytes and blastocysts cells derived from these oocytes (P < 0.05). The loss of ΔΨm in HS‐treated oocytes was not restored by exposure to BIP + BH4 and there was no effect in mtDNA copy number. In conclusion, the present results show that HS‐induced apoptosis in bovine oocytes involves Bax and BH4 domain‐dependent pathways. Mol. Reprod. Dev. 76: 637–646, 2009. © 2008 Wiley‐Liss, Inc.     

Determining the optimum temperature and salinity for larval culture,and describing a culture protocol for the conservation aquaculture for European smelt Osmerus eperlanus (L.)

Populations of anadromous European smelt Osmerus eperlanus (L.) are declining across its geographical range in northern Europe, but no practical culture techniques exist to develop stock enhancement programmes for this species. In this study, a culture protocol is described to rear fish from fertilised eggs to mature adults in 2 years involving the use of ‘green water’, live feed and artificial diets. The sequence of embryonic development for eggs incubated at 10°C/0 ppt was described and photographed. To determine the optimum conditions for larval culture, fertilised eggs were reared at a range of salinities (0–20 ppt) and temperatures (5–18°C) until first feeding. Best hatching success (ca. 97%), size at hatch (ca. 0.8 mm) and survival to first feeding (ca. 96%) of larvae were achieved under combined conditions of low salinity (0–0 ppt) and temperature (5–10°C). No larvae survived a salinity of 20 ppt. The time taken from fertilisation to hatch (FtH) and hatching duration (HD) were temperature-dependent ranging from 42 days FtH and 10 days HD at 5ºC, to 10 days FtH and 2 days HD at 18°C irrespective of salinity. The results indicate that conservation programmes could utilise existing salmonid hatchery facilities (i.e. freshwater, ≤10°C water temperature) for stock enhancement. Since on-growing of smelt involves the logistical and technical problems of live feed production, it is recommended that smelt enhancement programme utilise freshwater hatchery facilities to rear fish until hatching, and then stock out onto known spawning grounds in rivers allowing hatched larvae to drift into estuaries to complete the larval and juvenile phases. This approach would minimise the time spent in the hatchery post-hatching, eliminate the need for live food production, prevent the development of predator-naïve fish, and hence would mimic the natural life cycle of the species as closely as possible.     

Stressor interactions in freshwater habitats: Effects of cold water exposure and food limitation on early‐life growth and upper thermal tolerance in white sturgeon,Acipenser transmontanus

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Reproductive Biology of Three Commercially Important Hemiramphid Species in South-eastern Australia

Synopsis The reproductive biology of eastern sea garfish Hyporhamphus australis, eastern river garfish H. regularis ardelio, and snub-nosed garfish Arrhamphus sclerolepis were described throughout their respective ranges in the coastal waters of New South Wales (NSW), Australia. Peaks in gonadosomatic indices indicated that spawning of eastern sea garfish occurred in late spring and early summer (November–December) on the south coast of NSW, and in winter and early spring (June–September) on the north coast. Eastern river garfish spawned between July and December in NSW estuaries and snub-nosed garfish spawned between October and January in the Clarence River. The sex ratios in commercial catches of eastern sea garfish from the north coast of NSW were biased toward male fish, but approached equality for fish caught from the south coast. Sex ratios were significantly biased toward female snub-nosed garfish, and female eastern river garfish from all estuaries sampled. Mean (±SE) batch fecundity was 1498 ± 110 (range: 98 – 3449) ripe oocytes per female for eastern sea garfish, 917 ± 36 (range: 102 – 2268) ripe oocytes per female for eastern river garfish, and 716 ± 104 (range: 20 – 2956) ripe oocytes per female for snub-nosed garfish across the range of mature sizes examined. There was a linear relationship between batch fecundity and fish size for all three species of garfish. Eastern sea garfish approached 50% maturity at a larger size than snub-nosed, or eastern river garfish. Size at 50% maturity was similar for male and female eastern river and snub-nosed garfish, but male eastern sea garfish matured at a significantly smaller size than females. All three species appear capable of spawning in the spawning season immediately following the one in which they were born. Mature female fish of all three species had distributions of oocyte diameters consisting of three or four modes, which strongly suggests asynchronous oocyte development and a multiple spawning strategy during the spawning season. Implications for the management of garfish fisheries in NSW are also discussed.