Physical changes in specimens of five species of Cyprinidae preserved in ethanol and frozen

Preservation in 30% ethanol and freezing to a temperature of ?20 ± 2° C is an appropriate method for measurement of fish eggs, larvae and juveniles. Egg diameter of the common carp Cyprinus carpio increased insignificantly by 1·32% after preservation compared with live size. The total length (L_T) of 1 day post‐hatching (dph) larvae as well as the standard length (LS) of 16 dph larvae of C. carpio increased significantly (2·95 and 1·50%, respectively) after preservation. Egg diameter as well as the L_T of 1 dph larvae of barbel Barbus barbus increased significantly after preservation, by 1·74 and 1·96%, respectively over their original size. The standard length (L_S) of 14 dph larvae of B. barbus as well as juveniles of B. barbus, crucian carp Carassius carassius, common nase Chondrostoma nasus and tench Tinca tinca decreased significantly after preservation (?0·56 to ?5·54%), whereas their body mass increased significantly (11·46–18·57%). Preserved eggs of C. carpio and B. barbus were hard, round and transparent. The larvae and juveniles of examined fishes, preserved in frozen ethanol, were straight, flexible and easily measurable after 60 days. Integrity of body surface and fins, as well as preservation of colours were much better in larvae or juveniles frozen and thawed only once than in specimens frozen and thawed thrice. Cooling in 30% ethanol to a temperature of 6 ± 2° C and freezing in water to a temperature of ?20 ± 2° C are not appropriate preservation methods for eggs and larvae of C. carpio (1 and 16 dph).     

Ontogeny of turbiditaxis in hatchery‐reared Japanese anchovy Engraulis japonicus

The ontogenetic change in turbiditaxis (i.e. attraction to turbid waters) was examined in the larvae and juveniles of Japanese anchovy Engraulis japonicus by testing three levels of turbidity (0, 20 and 100 ppm of kaolin). Larvae of 12, 20 and 30 mm standard length (L_S) exhibited turbiditaxis to both 20 and 100 ppm of kaolin, whereas 6 mm L_S larvae and 45 mm L_S juveniles did not exhibit any turbiditaxis. Turbiditaxis might explain the ontogenetic habitat shift from coastal to offshore waters reported for this species.     

Effects of Temperature and Body Size on the Swimming Speed of Larval and Juvenile Atlantic Cod (Gadus Morhua): Implications for Individual-based Modelling

Synopsis The routine swimming speed (S) of three groups of 4, 9 and 32 cm total length (L_T) juvenile cod (Gadus morhua) was quantified in the laboratory at 6 – 10 different temperatures (T) between 3.2 and 16.7°C. At temperatures between 5 and 15°C, mean group S increased exponentially with increasing T (S=a e^bT) and the effect of temperature (b = 0.082, Q₁₀ = 2.27) was not significantly different among the groups (over the 8-fold difference in fish sizes of early- and post-settlement juveniles). Differences in mean S among individuals within each group were quite large (coefficient of variation = 40 – 80%). Swimming data for juveniles and those collected for groups of 0.4, 0.7 and 0.9 cm standard length (L_S) larvae were combined to assess the effect of body size on S. At 8°C, S (mm s⁻¹) increased with L_S (mm) according to: S = 0.26L_S^Φ−5.28L_S⁻¹, where Φ = 1.55L_S^−0.08. Relative S (body lengths s⁻¹) was related to L_S by a dome-shaped relationship having a maximum value (0.49 body lengths s⁻¹) at 18.5 – 19 mm L_S corresponding to the sizes of fish at the end of larval-juvenile metamorphosis. Previous larval cod IBM’s using a cruise-predator mode likely overestimated rates of foraging (prey searching and encounters) by a factor of ~2, whereas foraging rates in pause-travel models are closer to estimates of swimming velocities obtained in this and other laboratory studies.     

Captive hybridization of two geographically isolated pygmy angelfish species,Centropyge fisheri and Centropyge resplendens

This study documents the rearing of two pygmy angelfish species, Centropyge fisheri and Centropyge resplendens, and the early life history and reproduction of their hybrid offspring. A C. fisheri female, collected from Hawaii, and a C. resplendens male, captive‐bred from parental stock collected from Ascension Island, were maintained at the hatchery facility for 7 months. Continuous spawning was achieved at a photoperiod cycle of 14L:10D and a water temperature of 26·5° C, range ±1° C. Over the 110 day period, the C. fisheri female spawned 102 times, 57% of which resulted in embryos (fertilized eggs). The mean ±s.d. fecundity per spawn was 730 ± 459 eggs (range 52–1967). Fertility (% eggs that developed into embryos) of all eggs that were preserved was 22·4 ± 25·6%. A total of 235 hybrid juveniles were raised through metamorphosis with an average larval survival of 16·4%. Eight F₁ hybrid juveniles isolated for further study began to display signs of reproductive behaviour c. 300 days post‐hatch (dph). Spawn resulting in non‐fertile eggs were first obtained 319 dph, and fertilized eggs developing into embryos were obtained after 411 dph from at least two female individuals. While no attempt was made at rearing the F₂ larvae, embryo and larval development were normal up to 8 dph. Reproduction and development observed for all hybrid generations in this study were normal, similar to other Centropyge species and indicates a very close phylogenetic relationship between what are currently considered distinct species, e.g. C. fisheri and C. resplendens.     

Early growth and development of reciprocal hybrids of the starry flounder Platichthys stellatus and stone flounder Kareius bicoloratus

Larval growth and development of hybrid flounder were observed and compared with those of their parent species. The reciprocal hybrids of female starry flounder Platichthys stellatus and male stone flounder Kareius bicoloratus (hybrid Sb) and of female K. bicoloratus and male P. stellatus (hybrid Bs) both survived and grew to juveniles. Development was divided into nine stages (A–I). Many of the hybrids' traits were identical and intermediate to those of their parents. The position of the eye, however, was primarily sinistral in both hybrids (80% in Sb and 76% in Bs), a trait possessed by P. stellatus (80%) in the western Pacific Ocean. The daily growth rates of the larvae were similar. In the parent species, development was more rapid in P. stellatus than in K. bicoloratus whereas rate of development was intermediate in both Sb and Bs hybrids. The size at settlement [standard length (L_S) at stage H (mean ± s.d. )] was 9·82 ± 1·47 mm for the hybrid Sb and 9·99 ± 0·90 mm for the hybrid Bs, while the minimum age at metamorphosis (initial age at stage H) was 29 days after hatching (DAH) in both hybrids. In comparison, L_S at settlement in parent species was 6·43 ± 0·25 mm for P. stellatus and 12·87 ± 1·29 mm for K. bicoloratus. Minimum age at metamorphosis for the parents was 23 DAH at stage G in P. stellatus and 34 DAH at stage H in K. bicoloratus. Thus, the timing of settlement of hybrids was different from that of their parent species. These traits may occur with high frequency in a natural habitat.     

A finite interval of initial swimbladder inflation in Latris lineata revealed by sequential removal of water‐surface films

A finite interval of initial swimbladder inflation in striped trumpeter Latris lineata larvae occurred over 4 days at 16° C. Water‐surface films were removed on different days to form treatments: 4, 8, 9, 10, 11 and 12 days post hatching, dph (day 4, 8, 9, 10, 11 and 12 treatments, respectively). No swimbladder inflation was recorded prior to water‐surface film removal. When the water‐surface films were removed in day 4 and 8 treatments, initial swimbladder inflation was first recorded in larvae 9 dph at mean ± s .e . 35·0 ± 5·4%(n = 4) and 45·0 ± 7·9%, respectively. Water‐surface film removal at days 9, 10 and 11, resulted in initial swimbladder inflation the following day at 62·5 ± 2·5, 62·5 ± 7·2 and 11·3 ± 5·5% in larvae 10, 11 and 12 dph, respectively. No swimbladder inflation was recorded following water‐surface film removal on day 12. There was no significant difference in initial inflation among larvae in day 4, 8, 9 and 10 treatments, ranging from 65·0 ± 4·1 to 73·8 ± 6·9%(P > 0·05). Initial inflation was significantly lower in the day 11 treatment (11·3 ± 5·5%)(P < 0·05). During the inflation interval (9–12 dph) swimbladders displayed one of three morphologies; liquid dilation, gas inflated and collapsed. Collapse of the swimbladder lumen was first apparent in larvae without swimbladder inflation from 11 dph and progressively developed thereafter in all larvae with non‐inflated swimbladders. Larvae >6·1 mm standard length lost the ability to undergo initial swimbladder inflation. This study demonstrates that the interval for initial swimbladder inflation in striped trumpeter is short, finite and related to larval size. The end of the inflation interval was marked by onset of abnormal swimbladder morphologies, but not to closure of the pneumatic duct.     

Effects of feeding rate on the growth performance of gynogenetic albino sterlet,Acipenser ruthenus (Linnaeus, 1758) larvae

The feeding rate effects were studied on the growth performance of gynogenetic diploid larvae of sterlet Acipenser ruthenus during the first 4 weeks of exogenous feeding. The experimental rearing was conducted from 7 to 38 days post‐hatch (dph) in a circulation system. This was set up in four groups with three replicates (440 individuals/replicate), viz: AC‐control larvae fed Artemia sp., CFC‐control larvae fed compound feed, AG‐gynogenetic larvae fed Artemia sp., and CFG‐gynogenetic larvae fed compound feed. The larvae were reared in glass tanks (44 L volume, 10 individuals/L) with the temperature maintained at 18 ± 0.5°C, photoperiod of 12L:12D and water flow regime of 1‐L/min and fed 50%, 25%, 25%, and 9% of their total biomass/day during feeding. Highest TL and WBW of gynogenetic diploid larvae (AG) were observed with 50.6 ± 1.2 mm and 607.3 ± 36.1 mg (n = 30) at 38 dph. Highest TL and WBW of control larvae (AC) were recorded with 49.5 ± 3.8 mm and 600.8 ± 88.0 mg (n = 30), respectively, with 73.1% ± 11.4% survival; the lowest survival rate was at 46.4% ± 7.1% (n = 30) for the CFG group. The results indicate that the gynogenetic and normal larvae of sterlet fed with live food (Artemia nauplii) from 7 dph can achieve higher growth and survivability compared to the larvae fed with formulated test feed. Results of this study suggest that the effective rearing of sterlet larvae from 7 to 38 dph strongly depends upon the types of feed rather than the genome manipulation performed.     

Embryonic and larval traits of the temperate damselfish Chromis crusma reveal important similarities with other Pomacentridae throughout the family's thermal range

Embryonic development and larval morphology of Chromis crusma was described from five nests sampled between 21 and 25 m depth in central Chile (33°S). From each nest, a set of c. 100 randomly selected eggs were hand-collected and transported in seawater to the laboratory. Subsets of c. 30 eggs per nest were maintained in 50 ml glass containers at a constant ambient temperature of c. 12°C (range 11.5–12.9°C). Egg length (L) and width (W) and larval notochordal length (L_N) were measured from photographs. Geometric morphometric analyses were performed in newly hatched and 1 week old larvae to quantify shape changes. Ellipsoid eggs had an average (mean ± SE) size of 1.12 ± 0.05 mm L and 0.67 ± 0.02 mm W, with volume being similar throughout 15 developmental stages (i.e., ellipsoid-shaped; 0.27 mm³). Planktonic larvae hatched between 5 and 11 days at 12°C and had a mean L_N of 3.13 ± 0.25 mm, a yolk sack volume of 0.03 mm³ and an oil droplet volume of 0.005 mm³. Morphological traits at hatching included: (a) lack of paired fins and jaws; (b) single medial fin fold; (c) lack of eye pigmentation; (d) yolk sac present near anterior tip; (e) melanophores distributed along ventral surface with one pair over the forehead. In order to generate an up-to-date summary of developmental traits within Pomacentridae, we reviewed literature on egg development (e.g., shape and number of oil droplets), hatching and larval traits (e.g., morphology, pigmentation patterns). Thirty-two publications accounting for 35 species were selected, where eggs, embryonic development, hatching and larval traits were found for 26, 21, 24 and 34 species, respectively. In order to evaluate potential phylogenetic and environmental relationships within the early stages of Pomacentridae, cluster analyses (Bray Curtis similarity, group average) were also performed on egg and larval traits of 22 species divided by subfamily (Stegastinae, Chrominae, Abudefdufinae, Pomacentrinae) and thermal ranges (i.e., low: 16.5°C (range: 12–21°C), medium: 24.5°C (range:21–28°C) and high: 27°C (range: 26–28°C)), suggesting that early developmental patterns can be segregated by both temperature and phylogenetic relationships.     

Age,growth and hatch dates of ingressing larvae and surviving juveniles of Atlantic menhaden Brevoortia tyrannus

Ages, growth and hatch dates of ingressing Brevoortia tyrannus larvae were determined in a 3 year sampling survey at the mouth of the Chesapeake Bay, U.S.A. To determine if otolith‐aged cohorts had variable relative survival, hatch dates of summer‐caught young‐of‐the‐year (YOY) juveniles collected throughout the Chesapeake Bay were compared with hatch dates of ingressing larvae. Modal total length of ingressing larvae was similar among years: 28 mm in 2005–2006 and 2007–2008, and 30 mm in 2006–2007. Ages of ingressing larvae ranged from 9 to 96 days post hatch (dph); mean ages were similar among years, but significantly older in 2006–2007 (50 dph) than in 2005–2006 (44 dph) and 2007–2008 (46 dph). Larval growth rates differed among years. Earliest growth, when larvae were offshore (0–20 dph), was faster in 2006–2007 (0·62 mm day^?1), than in 2005–2006 and 2007–2008 (0·55 mm day^?1 in these years). Subsequently, from 30 to 80 dph, growth was slowest in 2006–2007. Hatch dates of ingressing larvae occurred from September to March and 90% (2007–2008) to 98% (2006–2007) had hatched prior to 31 December. In contrast, most surviving YOY juvenile B. tyrannus had hatched in January to February, suggesting selective mortality of early‐hatched individuals, apparently during the overwinter, larval to juvenile transition period.     

Influence of post-treatment temperature, solvents and emulsifiers on the toxicity of cypermethrin to larvae of Spodoptera litura (Fab.)

Larvae of the tobacco caterpillar, Spodoptera litura (Fab.) were topically bioassayed with cypermethrin emulsion formulation and proprietary EC in combination with different solvents and emulsifiers for their toxicity at 16 ± 2°C and 27 ± 1°C. Among different solvents, the descending order of efficacy (with relative toxicity in parenthesis) against 9 ± 1 day old larvae of S. litura (Fab.) at 16 ± 2°C than 27 ± 1°C was: xylene (7.41), aromex (5.54), cyclohexane (4.93), C-IX (3.93), benzene (2.40) and toluene (2.26). With emulsifier, the order was: Triton X-100 (7.41), SN₄S (4.60), Teepol (2.90), Tween-80 (1.49), Swascofix DP-50 (1.28), SN₄R (1.10) and S₅₂B (1.07). However, the proprietary cypermethrin EC was 2.31 times more toxic at 16 ± 2°C than at 27 ± 1°C.     

Growth and muscle cellularity of diploid and triploid Atlantic cod (Gadus morhua Linnaeus, 1758) larvae

The aim of this study was to compare somatic growth and muscle fibre development in diploid and triploid siblings of Atlantic cod (Gadus morhua Linnaeus, 1758) during the larval stage. Newly hatched larvae were transferred into 200‐L tanks, three tanks per ploidy group (70 larvae L^?1, continuous light, gradually increasing seawater temperature 7–11°C and flow rates 50–117 L h^?1). Larvae were fed rotifers from 2 to 22 days post hatch (dph), Artemia 19–31 dph and weaned onto a microparticulate diet from 26 dph until the end of the experiment. Measurements of growth (dry weight, standard length) and muscle cellullarity were taken at intervals between 1 and 44 dph. Ploidy groups showed a similar performance throughout the trial, although a marked stagnation in growth was observed for triploids during the weaning from Artemia onto dry feed. Overall, diploid and triploid cod larvae showed a similar development in muscle fibre growth pattern during the experimental period. For both groups, the total number of fast muscle fibres showed a 10‐fold increase (from 384 to 3462), whereas the diameter of fast fibre increased from 8.9 to 13.3 μm (mean number from all treatments). Thus, a temporary but significant effect of triploidy on fast muscle fibre growth pattern was observed in 19 dph larvae in terms of fibre size and number, with triploids showing larger mean fast fibre diameter (11.62 ± 0.63 vs. 10.05 ± 0.34) and a lower number of fibres with a diameter <5 μm than their diploid siblings. Thus, this was found to be related to larvae size and to the differences in total fast fibre cross sectional areas rather than to ploidy status. Overall, our results suggest possible deficiencies in nutrients’ digestion and absorption of triploid cod larvae particularly during the transitional period from live food to inert diets.     

Acute Toxicity of Arsenic under Different Temperatures and Salinity Conditions on the White Shrimp Litopenaeus vannamei

The aim of this study was to determine acute toxicity in the post larvae of the white shrimp Litopenaeus vannamei after 96 h of exposure to dissolved arsenic under three different temperatures and salinity conditions. Recent reports have shown an increase in the presence of this metalloid in coastal waters, estuaries, and lagoons along the Mexican coast. The white shrimp stands out for its adaptability to temperature and salinity changes and for being the main product for many commercial fisheries; it has the highest volume of oceanic capture and production in Mexican shrimp farms. Lethal concentrations (LC₅₀–96 h) were obtained at nine different combinations (3?×?3 combinations in total) of temperature (20, 25, and 30 °C) and salinity (17, 25, and 33) showing mean LC₅₀–96 h values (±standard error) of 9.13?±?0.76, 9.17?±?0.56, and 6.23?±?0.57 mgAs?L^?1(at 20 °C and 17, 25, and 33 salinity); 12.29?±?2.09, 8.70?±?0.82, and 8.03?±?0.59 mgAs?L^?1 (at 25 °C and 17, 25, and 33 salinity); and 7.84?±?1.30, 8.49?±?1.40, and 7.54?±?0.51 mgAs?L^?1 (at 30 °C and 17, 25, and 33 salinity), respectively. No significant differences were observed for the optimal temperature and isosmotic point of maintenance (25 °C–S 25) for the species, with respect to the other experimental conditions tested, except for at 20 °C–S 33, which was the most toxic. Toxicity under 20 °C–S 33 conditions was also higher than 25 °C–S 17 and 20 °C (S 17 or 25). The least toxic condition was 25 °C–S 17. All this suggests that the toxic effect of arsenic is not affected by temperature changes; it depends on the osmoregulatory pattern developed by the shrimp, either hyperosmotic at low salinity or hiposmotic at high salinity, as observed at least on the extreme salinity conditions here tested (17 and 33). However, further studies testing salinities near the isosmotic point (between 20 and 30 salinities) are needed to clarify these mechanisms.     

Induction of ovarian maturation and development of eggs,Larvae and Juveniles of the Puffer,Takifugu exascurus,Reared in the Laboratory

Ovarian maturation of the pufferTakifugu exascurus (Jordan et Snyder) was induced, and embryonic, larval and juvenile development was observed. The brood fish were collected in Tassha Bay, Sado Island (38°05′N, 138°15′E), during the spawning season in 1986 which seemed to extend from late June to mid-July. To each female 3 mg acetone-dried pituitary ofHypophthalmichthys molitrix was injected to induce ovarian maturation, which took place in about 77 hours at a water temperature of 19.5–21.0°C. The eggs obtained by hormone injection were artificially fertilized with the milt from a collected male. The hatched larvae were fed successively on rotifersBrachionus plicatilis, Artemia nauplii and minced fish meat, and reared for a period of about one year. The eggs were spherical, 1.24±0.04mm in diameter, demersal and adhesive. The egg-membrane Was transparent and yolk was orange in color, containing a cluster of small oil-globules. The incubation period was about 160 hours at a water temperature of 18.5–21.0°C. The newly-hatched larvae, measuring 2.9– 3.1 mm TL, had 8+15 = 23 myomeres. Absorption of the yolk was completed 3 days after hatching, by which time the larvae had attained 3.5–3.6 mm TL. Larval finfolds disappeared and rudimentary dorsal, anal and caudal fins formed at 4.1–4.4 mm TL, in 6 days after hatching. In 9-day old larva (5.4 mm TL), fin ray rudiments appeared on the dosai, anal and caudal fins and spine-like scale formed on the belly. In 16-day old specimens, 9.1–10.2 mm TL, the full complements of fin rays were completed on all the fins and the fish reached the juvenile stage. The growth of larvae and juveniles reared in 1986–1987 is expressed by the following equations, where y is total length (mm) and x is days after hatching. y₁ = 2.9420· 1.0639^x 0 ≦ x ≦ 19 (r = 0.998) y₂ = 4.0286· 1.0464^x 19≦ x ≦ 33 (r = 0.998) y₃ = 9.8854· 1.0180^x 33 ≦ x ≦ 72 (r = 0.996) y₄ = 20.1555· 1.0080^x 72 ≦ x ≦ 115 (r = 0.998) y₅ = 28.0610· 1.0049^x 115 ≦ x ≦ 202 (r = 0.995)     

Occurrence of Gobiidae larvae in a tropical Brazilian estuary,with particular emphasis on the use of size classes to categorize species guilds

The structure and seasonal dynamics of larvae of the Gobiidae family in the Mucuri Estuary (Bahia, Brazil) were studied for nine consecutive years. Sampling was conducted at three stations in the lower estuary channel, between 2002 and 2010, in relation to season, day and night and tidal variations. A total of 5802 Gobiidae larvae, representing 15 taxa (12 species and three morphotypes), were collected in the Mucuri Estuary during this time. The highest mean ± s.d. density of fish larvae, 54·7 ± 79·8 larvae 100 m^?3, was recorded during the flood tide and night sampling. Ctenogobius boleosoma was the most abundant species (68%), being dominant in the rainy and dry seasons and had a long reproductive period. This species was classed as a marine estuarine‐opportunist because it was observed at high frequencies and active larvae entering the estuary between 6·1 and 12·0 mm standard length (L_S). Gobionellus oceanicus, second in abundance (12%), occurred only in later larval stages but did not use the estuary as a preferred location for spawning, being classed a marine estuarine dependent. Microgobius carri (11%) was represented in all L_S classes and was resident in the estuary for spawning, remaining there throughout their life cycle. The other species were considered rare due to their low densities and could not be classified in any guild.     

Implication of storage conditions on luminescence response from Photobacterium phosphoreum in free and immobilized form

Bioluminescent bacteria in the form of a cell suspension for on-site hazard analysis are not suitable as in vivo luminescence in free cells fluctuates and may lead to erroneous results. Furthermore, the culture broth cannot be stored for long durations to continue sensing analytes as the luminescence ceases over time. Factors that affect luminescence response include growth dynamism, and ambient environmental conditions. The present study investigated the effect of storage conditions such as temperature (25 ± 2°C, room temperature; 4°C; and −20°C) and ambient aqueous environment (M1: sucrose, 1.02 M; M2, bioluminescent media [tryptone, 10 g L⁻¹; NaCl, 28.5 g L⁻¹; MgCl₂.7H₂O, 4.5 g L⁻¹; CaCl₂, 0.5 g L⁻¹; KCl 0.5 g L⁻¹; yeast extract, 1 g L⁻¹; H₂O, 1 L]; M3, bioluminescent media and 95% glycerol, 1:1 ratio) on the luminescence emission from the calcium alginate-immobilized Photobacterium phosphoreum (S_b) against the cells in free suspension for an extended period. The results indicated that both the parameters that were undertaken markedly affected the luminescence. In the study, S_b showed an enhanced luminescence emission than the control up to 18.5-fold and for a prolonged period which can be efficiently utilized for rapid biosensing of hazardous materials.     

Survival of Heleomyza borealis (Diptera, Heleomyzidae) larvae down to − 60°C

Heleomyza borealis Boh. (Diptera, Heleomyzidae) overwinters as larvae in Arctic habitats, where they may experience winter temperatures below ? 15°C. The larvae freeze at c.? 7°C but in acclimation experiments 80% survived when exposed to ? 60°C. Of the larvae exposed to between ? 4 and ? 15°C, only 3% pupated. However, when cooled to ? 20°C this increased to 44%, with 4% emerging as adults. Larvae maintained at 5°C contained low levels of glycerol, sorbitol and trehalose, which did not increase with acclimation to low temperatures. However, levels of fructose increased from 6.1 μg mg^?1 fw in control animals to 17 μg mg^?1 fw when exposed to ? 2°C for 1 week. Larval body water (2.2 ± 0.1 g/g dw, mean ± SD, n = 100) and lipid content (0.22 ± 0.002 g/g fw, mean ± SE) showed no significant change during acclimation to low temperatures. Larvae maintained at a constant 5°C survived for over 18 months with little loss of body mass (from 7.5 ± 1.2 to 7.0 ± 1.2 mg fw, mean ± SD, n = 20), but none pupated. Heleomyza borealis larvae appear to feed and grow until they reach a body mass of about 7.5 mg and then become dormant. They remain in this state until they experience a low temperature stimulus (< ? 15°C) followed by a warm period (≈ 5°C). This ensures that the larvae pupate and adults emerge in early summer, allowing the maximum growing period before the following winter. Heleomyza borealis are adapted to survive long winters in a dormant larval state. They have a low metabolic rate, can conserve body water even at subzero temperatures but do not synthesize large quantities of cryoprotectants.     

Sex steroid changes during temperature‐induced gonadal differentiation in Paralichthys olivaceus (Temminck & Schegel, 1846)

Sex steroid changes during temperature‐induced gonadal differentiation were evaluated in the olive flounder, Paralichthys olivaceus. Larvae were reared at 21 ± 0.5°C, 24 ± 0.5°C and 28 ± 0.5°C from day 40 post‐hatching (dph) to 90 dph. The proportion of males was 61.1, 76.7, 87.8 and 47.8% in 21°C, 24°C, 28°C and in control groups, respectively. Gonadal differentiation was circa 65 dph, when fishes were a mean 39 mm total length (TL). The gonads developed faster when fishes were reared in higher temperatures. Radioimmunoassay (RIA) analyses indicated that the level of estradiol‐17β (E2) changed during the period of gonadal differentiation and peaked at an onset of ovarian differentiation in all groups. Compared with fish in control groups, the levels of E2 were lower in thermal‐treated groups, especially in the highest temperature groups. The present results indicate that E2 plays a major role in the process of ovarian differentiation, and suggest that temperature‐induced masculinization in P. olivaceus is mainly due to a decrease in the E2 level during the period of ovarian differentiation.     

Some effects of freezing temperatures on overwintering larvae of the large elm bark beetle (Scolytus scolytus)

Overwintering final instar larvae of Scolytus scolytus were tested for coldhardiness by exposure to a range of sub-zero temperatures (–7°Cto –31 °C) in a frost-gradient apparatus for 7 days. The Lt₅₀ for larvae removed from the bark (– 20.5 °C) was significantly different (P < 0.01) from the Lt50 (–18.3 °C) for larvae insulated by the bark (thickness of 7 mm ± 2 mm). Larvae with food in their digestive tract were more susceptible to freezing than the overwintering final instars which had voided their stomach contents. Duration and intensity of cold did not cause any adverse long-term effects. Most of the larvae which survived the sub-zero treatments were able to pupate or reach the adult stage. The mean supercooling point of the overwintering larvae (–30.85 °C) confirmed their cold-hardiness.