Autotriploid tench Tinca tinca (L.) larvae obtained by fertilization of eggs previously subjected to postovulatory ageing in vitro and in vivo

Eggs of diploid tench Tinca tinca were half-stripped out and stored for 0 (control batch), 1, 3 and 5 h at mean ± s . d . 17·0 ± 0·4 and 21·9 ± 0·5° C or for 0, 1, 2, 3, 4 and 5 h at 24·0 ± 0·0° C in vitro prior to fertilization. The eggs remaining in vivo in the fish kept at 17·0 ± 0·4 and 21·9 ± 0·5° C were collected and fertilized in the same time intervals. Fertilization rate and larval yield mostly decreased after 3–5 h storage of eggs both in vitro and in vivo and only the diploid larvae were found in all control batches. Triploid larval yields increased to a maximum 5·26% after 5 h in vitro storage at 24·0° C and 1·07 and 1·60% after 3 h in vitro storage at 21·9 and 17·0° C, respectively. Triploid larval yield during in vivo storage at 21·9° C reached a maximum 0·91% after 5 h. As the spontaneous autotriploid larvae arose solely from fertilized eggs previously subjected to postovulatory egg ageing by means of prolongated storage, the autotriploidy was probably caused by failure of extrusion of the second polar body.     

Captive breeding and embryonic development of Chiloscyllium punctatum Muller & Henle, 1838 (Elasmobranchii: Hemiscyllidae)

The fecundity, embryonic development, growth and viability of captive Chiloscyllium punctatum were investigated over two breeding and laying seasons. Chiloscyllium punctatum is an oviparous species, which lays rectangular eggs from late July to February. The six adult female C. punctatum in the open ocean display tank at UnderWater World, Mooloolaba, Australia laid a total of 692 eggs between July 2004 and February 2006, with 228 of those being viable. This equates to each female laying 115·3 eggs over the two seasons, with 38 viable. The embryos are first visible to the naked eye at c . 18 days post deposition (dpd), while pre-hatching is first seen c . 35 dpd, and the embryos hatch at an average of 153 dpd (temperature 21–25° C). Embryonic growth is most rapid from 99 dpd until hatching. An average of 21·4% of the eggs hatched over the two seasons.     

Fertilization in landlocked sea lamprey: storage of gametes, optimal sperm: egg ratio, and methods of assessing fertilization success

Ovulated, unfertilized eggs of sea lamprey Petromyzon marinus could be stored for 1 day at 15° C without significant loss of fertilizing ability. After 2 days storage most eggs could still be fertilized. Lamprey semen could be stored up to 1 day. Thereafter, a decrease in sperm fertilizing ability occurred, accompanied with a decrease in sperm motility. Unlike teleost fish, sea lamprey eggs could still be fertilized after 1 h contact with water. This extended time of gamete fertility after release into water may help to account for the reproductive success of this species. Maximal fertilization rates were obtained at a sperm: egg ratio of 50 000, a ratio recommended for studies on fertility of individual males. Assessing fertilization success 3 min after fertilization (at cytoplasmic bleb stage) or 5 h after fertilization (at two–cell embryo) was strongly correlated ( r =0·92 and 0·98) with estimation and fertilization success at hatching. These results offer improvement in artificial fertilization techniques under laboratory conditions and provide new information on the biology of fertilization in sea lamprey.     

Holding temperature during the breeding season influences final maturation and egg quality in common wolffish

Mature common wolffish, Anarhichas lupus L., previously held at 8°C for several months, were exposed to 4, 8 or 12°C throughout the breeding season (November-January). Regardless of temperature, some females ovulated in early-November, but thereafter ovulation was delayed in the 8 and 12°C groups compared with the 4°C group. Temperature experienced by the adult fish also affected egg development and survival even though eggs were incubated at a common temperature of 6°C. The percentages of normally cleaved eggs (at the 32-cell stage) and egg survival to the eyed stage were significantly lower in the 12°C group (8%) than in the 4°C (73%) and 8°C (48%) groups. The differences in egg survival were also significant between the 4 and 8°C groups. Egg survival was also influenced by the length of time the broodfish had been exposed to the different temperatures: at 4 and 12°C there were significant positive and negative relationships, respectively, between egg survival and the number of days for which the female had been exposed to the given temperature prior to ovulation. Temperature experienced by the female did not affect egg size, relative egg weight, fertilization rate, relative fecundity or the number of ovulating individuals. The results indicate that both the timing of ovulation and egg quality of common wolffish are affected by the temperature experienced during the breeding season.     

Early development of the skull of Sander lucioperca (L.) (Teleostei: Percidae) relating to growth and mortality

The early osteological development of the skull (chondrocranium and osteocranium) of the pikeperch Sander lucioperca was studied. Specimens were reared at two temperatures, 15·5 and 18·0° C, from hatching until 47 and 43 days after fertilization (DAF), respectively. The skeletal elements characteristic for the different developmental stages were the same at both rearing temperatures, but pikeperch reared at 15·5° C reached the developmental stages later. The formation of the functional complexes, the neurocranium, jaws and suspensorium, branchial basket and hyoid arch, was evaluated chronologically. The focus was on skull development during several functional changes: at hatching, at the shift from endogenous to mixed feeding, the shift to exclusively exogenous feeding and upon reaching the final prey-capture mechanism. Growth in total length differed between fishes reared at the two temperatures, except during a phase of very slow growth from the end of the embryonic stage until the second larval stage. The latter phase, in which most of the bony elements of the viscerocranium started to form, was marked by high mortality. When exogenous feeding began, the growth rates at both temperatures increased distinctly and the first bony elements were formed in the neurocranium. Specimens reared at 18·0° C grew continuously, but those at 15·5° C showed a second period of slow growth and high mortality. Fish reared at 18·0° C reached the successive larval stages distinctly earlier than fish reared at 15·5° C.     

Reproductive biology of the endangered Oxleyan pygmy perch Nannoperca oxleyana Whitley

The reproductive biology of the Oxleyan pygmy perch Nannoperca oxleyana is described from simultaneous studies of wild populations in north-eastern New South Wales and mature fish held in aquaria. In the wild, 50% of males and females matured at total lengths of 24·0–25·9 and 28·0–29·9 mm, respectively. The species displays sexual dichromatism during the spawning season, with males developing more intense red and brown fin and body colouration, and black pelvic fins. Captive male broodfish displayed territoriality during the breeding season, closely guarding sites within artificial, plant-like substrata in which pairs of fish spawned adhesive eggs. Protracted serial spawning of wild and captive fish occurred from September to April and May at mean water temperatures ≥16·6° C and day length ≥10·7 h. Captive broodfish spawned on an average of 57% of days during the 256 day spawning period. Gonado-somatic indices averaged 0·7% for all ripe males and 4·1–4·2% for all ripe females collected. Mean total and batch fecundities of captive females were 1323 eggs per fish and 7·8 eggs per fish per day, respectively, and relative fecundity was 587 eggs g⁻¹ of body mass. Batch fecundity of wild females was estimated at 7·8 eggs per fish. The adaptive significance of this reproductive strategy in a harsh, variable environment is discussed.     

Cry (preservation of spermatozoa of the whitefish Coregonus muksun Pallas

The cryopreservation methods for whitefish, Coregonus muksun , spermatozoa were studied using six different extenders with -40°C and -196°C as storing temperatures. In 1980, the spermatozoa of individual fishes were frozen in dry ice, in vials containing three different media based on glycerol as cryoprotectant. The pure glycerol media resulted in a 20% average rate of fertilization when samples were stored at — 40°C. The thawing procedure did not affect fertilization. In 1981, the sperm of individual fishes was frozen in pellets and stored in liquid nitrogen. Cryopreserved semen thawed in 0·119M NaHCO₃ produced 29·6–87·7% eyed eggs. The differences were caused mainly by the type of extender. The highest rate of fertility was obtained using an extender consisting of 0·3 M glucose with 20% glycerol (87·7% eyed eggs; 98·6% eyed eggs when compared to the corresponding control). The same values, using the modified extender of Stoss (Stoss & Refstie, in press), were 38·7–82·4% and, on the average, 90% eyed eggs, respectively.     

Hypoxia, low salinity and lowered temperature reduce embryo survival and hatch rates in black bream Acanthopagrus butcheri (Munro, 1949)

Embryo survival and hatch rates were measured in black bream Acanthopagrus butcheri exposed to different treatments of dissolved oxygen: hypoxic and normoxic, three salinities: 15, 23 and 35 and two temperatures: 16 and 20° C. Hypoxic conditions (50% saturation) reduced 1 day embryo survival by up to 15% ( P < 0·05) compared to embryos held in normoxic (>80% saturation) conditions. Temperature had no effect on the survival of embryos in these treatments, however, lowered salinity significantly reduced embryo survival at 20° C, but not at 16° C. Mean hatch rates were reduced by 10–28% in hypoxic treatments ( P < 0·05) and lowered salinity treatments ( P < 0·05). Hatching was delayed by up to 24 h at 16° C and very low (or zero) hatching occurred in hypoxic treatments at salinities of 15 and 23. These results confirm that environmental conditions in estuaries are important factors in determining spawning success of black bream and are discussed in relation to global warming and climate change that are likely to alter the physical conditions in southern Australian estuaries.     

The influence of temperature on the stage at hatch of laboratory reared Gadus morhua and implications for comparisons of length and morphology

A staging table for yolk‐sac larvae of Atlantic cod Gadus morhua was developed and applied to determine the development stage at hatch. Larvae ( n  = 20) were sampled during the period from 50% hatch to first‐feeding from four temperature treatments ( c . 2, 4, 8 and 12° C). Data recorded included standard length, development stage, yolk‐sac area and evidence of feeding by larvae. The results indicated that the stage of development at hatch was positively correlated to larval size. Furthermore, developmentally based changes in morphology shortly after hatch reduced much of the initial difference in size, although some difference still existed. No relationship between size at hatch and the incidence of first‐feeding larvae was found. Based on these observations, it is recommended that the development stage of hatching larvae be reported in studies investigating size and morphological differences when observing individual larval fish that originate from different populations, batches or species.     

Hypoxia tolerance in Atlantic cod

Oxygen saturation levels that killed 50 and 5% of cod Gadus morhua over 96 h averaged 21·2 and 27·7%, respectively. No fish survived at 10% saturation and only a few survived at 16% saturation, whereas no mortality occurred at 34 and 40% oxygen saturation. Since metabolic rate and oxygen consumption increase with increasing temperature, we hypothesized that cod would be less tolerant to hypoxic conditions at 6 than at 2° C. However, temperature (2 and 6° C) had no measurable impact on cod survival. Small (mean & S.D.; 45·2 ± 4·2 cm) and large (57·5 ± 3·8 cm) cod had the same tolerance to hypoxia. At the end of the experiments, hypoxia had a significant effect on blood haematocrit, mean cellular haemoglobin content, liver lactate, plasma glucose and plasma lactate, but accounted for only a small fraction (< 10%) of the variation, except for plasma lactate which exhibited a strong response with concentrations increasing progressively with decreasing levels of oxygen saturation. Temperature had a significant effect on most variates in normoxia and hypoxia. Variates also affected by oxygen level showed significant interactions between oxygen and size or temperature effects. However, these interactions accounted for only a small proportion of the variation. Physiological parameters indicated that extending the duration of our tests beyond 96 h would not have changed our estimates of the lethal thresholds. Hypoxic conditions are a permanent feature of the deep waters of the Gulf of St Lawrence. This study shows that a significant portion of the benthic habitats in the Gulf are uninhabitable for cod which would be expected to avoid waters below 28% oxygen saturation.     

Effects of temperature on developmental performance, survival and growth of sea lamprey embryos

This study assesses the influence of thermal regime on the development, survival rates and early growth of embryos of sea lamprey Petromyzon marinus incubated at five constant temperatures (7, 11, 15, 19 and 23° C). The time from fertilization to 50% hatching and from hatching to 50% burrowing were inversely related to incubation temperature. All the embryos incubated at 7° C died at very early stages, while those maintained at 11° C did not attain the burrowing stage. Survival from fertilization to hatching was 61, 89, 91 and 89% at 11, 15, 19 and 23° C, decreasing to 58, 70 and 70% from hatching to burrowing at 15, 19 and 23° C, respectively. Larvae reared during the first 3 months of exogenous feeding in a common environment at constant 21° C, revealed maximum survival for an incubation temperature of 15° C (43% of burrowed larvae) decreasing strongly at 19° C (16%) and 23° C (one suvivor among 240 larvae). Body length at the burrowing stage was maximum for embryos incubated at 19° C, but body mass increased in the interval 15–23° C. Mean incubation temperatures experienced by 117 broods during the embryonic development in the source river were estimated in 15·3±2·30° C and 16·7±1·76° C (mean±1 s.d .) for the periods fertilization-to-hatching and hatching-to burrowing, respectively.     

Effects of essential oil from mint (Mentha piperita) on Salmonella enteritidis and Listeria monocytogenes in model food systems at 4° and 10°C

The effect of mint ( Mentha piperita ) essential oil (0·5, 1·0, 1·5 and 2·0%, v/w) on Salmonella enteritidis and Listeria monocytogenes in a culture medium and three model foods; tzatziki (pH 4·5), taramosalata (pH 5·0) and pâté (pH 6·8), inoculated at 10⁷ cfu g^-1, at 4° and 10°C for ca 1 week was studied. In the culture medium supplemented with the essential oil, no growth was observed over 2 d at 30°C determined by a conductance method with a Malthus 2000 growth analyser. Salmonella enteritidis died in tzatziki in all treatments and declined in the other foods except for pâté at 10°C as judged with viable counts. Listeria monocytogenes populations showed a declining trend towards the end of the storage period but was increased in pâté. Mint essential oil antibacterial action depended mainly on its concentration, food pH, composition, storage temperature and the nature of the micro-organism.     

Geographical variation in egg cold hardiness: a study on the adaptation strategies of the migratory locust Locusta migratoria L.

Abstract. 1. For many species of insect, cold hardiness is an important trait that enables a population to develop in the next season and to extend its range. To elucidate the role of cold hardiness of the migratory locust Locusta migratoria L. in its outbreak and distribution areas, egg cold hardiness was examined in locusts derived from four locations from latitude 18°23'N to latitude 41°10'N in eastern China.
2. The supercooling points of eggs from different geographic populations did not differ significantly for the first development stage, with an average ± SE of −24.5 ± 0.51 °C, or for the second stage, −22.06 ± 0.68 °C, however there was a significant difference for the embryonic development phase among the four geographical populations. The egg supercooling point increased gradually from neonatal egg to old egg; eggs prior to hatching always had a much higher supercooling point.
3. Comparisons of the cold hardiness of four populations were carried out by validating the close correlation between latitude and the effects of cold on hatching, low lethal temperature (Ltemp₅₀), and low lethal time (Ltime₅₀). There were significant differences among the four populations; the northern population was more cold hardy than the southern population, and the two mid-latitude populations were intermediately cold hardy.
4. The cold hardiness of all populations was enhanced to various degrees by short-term cold acclimation at 0 °C and 5 °C. For most populations, a 2-day acclimation period seemed to be optimal.     

Temperature effects on embryonic development and the life cycle of Diphyllobothrium dendriticum

Diphyllobothrium dendriticum eggs collected from hamster faeces were incubated at 10 or 20 degrees C, or maintained at 4 degrees C for 11-30 months. On day 65, 20-50% of eggs failed to hatch at 10 degrees C and 42-51% did not hatch by day 21 at 20 degrees C. Our study indicates that eggs begin to hatch in mid-August and persist until October in many lakes within the natural range of D. dendriticum. Our results demonstrate that eggs stored at 4 degrees C will hatch, suggesting they can persist in the environment for long periods and contribute to the D. dendriticum life cycle in high Arctic lakes by hatching months or years after release.     

Thermal effects on growth and time to starvation during the yolk-sac larval period of Atlantic mackerel Scomber scombrus L.

The effect of incubation temperature (8·6, 11·1, 13·2, 15·1 and 16·8° C) on north-east Atlantic mackerel Scomber scombrus development, growth and age at starvation during the yolk-sac larval period was investigated. Standard length at hatch was found to be inversely proportional to incubation temperatures within the natural thermal ranges of this species; it ranged from 3·76 mm at 11·1° C to 3·30 mm at 17·8° C. Following hatch, however, larval growth rate was positively related to temperature. Individual logistic models, as a function of temperature and age, were fitted to the development processes of gape, eye pigmentation, jaw mobility and yolk exhaustion. Thereafter, development was classified into different ordered stages and an extended continuation model was fitted to the multinomial ordered stage classification. In all cases, there was a difference of >23 h between the first and the last individual developing in certain stage. The probability of survival decreased with age and was inversely related to temperature. Yolk utilization varied from 4·5 to 8·6 days and individuals died between 7·9 and 12·2 days from 17·8 to 11·1° C. The study demonstrated the significant impact that temperature has on development, growth and survival rates, during the early life history.     

Geographic parthenogenesis in the stream mayfly Eurylophella funeralis in eastern North America

Unisexual populations were found largely at the periphery of the geographic range of the mayfly Eurylophella funeralis in eastern North America. Bisexual populations generally had a normal sex ratio but at least two populations were observed with only about 2–11% males. Hatch success for unfertilized eggs depended on temperature but in general averaged >61% in unisexual populations and < 14% in bisexual populations. Eggs took about 65 d to hatch at 10°C and 19 d at 25°C; no hatching occurred at 5 or 30°C. Successful oviposition was observed for about 97% and 20% of unmated female adults from unisexual and bisexual populations respectively. The hatch success of unfertilized eggs for three geographically distinct bisexual populations was in versely correlated with the proportion of males in the populations. The genetic structure of bisexual and unisexual populations was very similar. Unisexual populations consists of clones, with as few as four recognizable phenotypes being present at a given location. All offspring had the same phenotype as the mother. Tychoparthenogenesis is suggested as the most reasonable working hypothesis concerning the origin of unisexual populations in E. funeralis .     

Temperature-induced changes of survival, development and yolk partitioning in Chondrostoma nasus

Fertilized Chondrostoma nasus eggs were incubated at 10, 13, 16 and 19° C until full resorption of the yolk sac. High survival was observed at 10–16° C (89–92% at the onset of external feeding), whereas at 19) C survival was depressed (76%). The time at which 5, 50 and 95% of individuals had hatched, filled the swim bladder, ingested the first food and fully resorbed the yolk sac was determined. An increase in temperature accelerated development and made it more synchronous. Within the period from fertilization to hatching embryonic development was theoretically arrested (t_{0 dev}) at 8·8° C, and growth was arrested (t_0gr) at 8·86° C. For the whole endogenous feeding period (from fertilization to full yolk resorption) the amount of matter transformed into tissue was temperature independent between 10° and 19° C. Respiration increased exponentially with age; the respiration increase was faster at higher temperatures, but, in general, metabolic expenditures of C. nasus were low. As a consequence, the efficiency of utilizing yolk energy for growth was high as compared with other fish species (57% during the whole endogenous feeding period); it was temperature independent. However, time was used less efficiently at low temperatures, increasing a risk of predation. Within the endogenous feeding period a shift from lower to higher temperatures for optimal yolk utilization efficiency was observed. The temperatures optimal for survival and energetic performance seem to be 13–16° C for egg incubation and 15–18° C for rearing of yolk-feeding larvae. Chondrostoma nasus is a potential candidate for aquaculture for restocking purposes.     

Influence of egg vitamin A status and egg incubation temperature on subsequent development of the early vertebral column in Atlantic salmon fry

The effect of egg vitamin A (VA) status and egg incubation temperature on the development of spinal disorders was investigated in Atlantic salmon Salmo salar fry. Atlantic salmon eggs were sorted into two groups with high VA (3·3 ± 0·1 μg retinol g⁻¹ dry mass) and low VA (2·2 ± 0·3 μg retinol g⁻¹ dry mass) status before fertilization and incubated at high (14° C) or low (8° C) temperature from 133 day degrees until the onset of feeding. High egg incubation temperatures increased the concentration of retinol in the eggs: the high VA and high temperature group displayed a significantly higher retinol concentration than the high VA and low temperature group ( P  = 0·001). After hatching, all experimental groups increased their retinol concentration. The source of the increased retinol levels was probably retinal, although astaxanthin may also be a VA precursor after hatching. Atlantic salmon fry incubated at high temperatures had increased amounts of notochord tissue. When measuring morphogenic activity in the notochord using the expression of sonic hedgehog ( shh , mRNA), however, no significant difference was found between the experimental groups. No clear effect of VA status or incubation temperature could be found on the formation of the early vertebral column although Atlantic salmon fry incubated at low temperatures had less regular constrictions of the prospective vertebral column than fry incubated at high temperatures.     

Effect of the ethanol content of beer on the heat resistance of a spoilage Lactobacillus

D -values of a heterofermentative beer spoilage lactobacillus were measured at 55°C, 60°C and 65°C in beers containing <0·05% to 4·4% v/v ethanol. Z -values for the different beers varied between 9·17 and 12·13°C. At each temperature an increase in ethanol reduced the measured D -value. The maximum, 5·01 min was observed in alcohol-free beer (<0·05%) at 55°C and the minimum, 0·31 min, at 60°C and 65°C in beer containing 4·4% ethanol. D -values could be increased by prior growth in the presence of ethanol. They could be reduced by adding ethanol to alcohol-free beer or by increasing its hop extract content. The implications for the pasteurization of low-alcohol beers are discussed.