To hatch or not to hatch? Egg hatch response to larval density and to larval contact in a treehole mosquito

Abstract.

• 1 We investigated the effect on egg hatch of exposure to: (1) varying larval density, and (2) larval contact in Aedes triseriatus Say (Diptera: Culicidae). For 2 days in the laboratory we submerged eggs into a treehole water medium containing 0 (control), 4, 12 or 24 larvae that could either contact the eggs directly or were separated from them by a screen. Following treatment, abundance of microorganisms on the egg surfaces, a food source for newly hatched larvae and a proposed hatching stimulus, was assessed by counts made from serial dilutions of samples.
• 2 We discovered a complex hatching response to larval contact and to larval density, and an interaction between these two factors in their effect on microbial growth. Hatching was inhibited in the 0-larva control, even though microorganisms grew abundantly on the eggs. Hatch rate, as well as microbial counts, were high for eggs in direct contact with 4 larvae. As density increased in the larval contact treatment, microorganisms disappeared from the egg surfaces and hatch rate declined.
• 3 When protected from larval grazing, eggs supported numerous microbial colonies irrespective of larval density. In contrast to the contact treatment group, egg hatch increased with increasing larval density. These observations suggest that the combination of microbial growth and a larval factor stimulates hatch. This hatching response may have evolved because both abundant microorganisms and numerous larvae reflect a habitat of good quality.

     

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.     

Performance of a spring-feeding moth in relation to time of oviposition and bud-burst phenology of different host species

Abstract. 1. As a spring-feeding moth committed to immature foliage, the autumnal moth Epirrita autumnata (Lepidoptera, Geometridae) must have egg hatch synchronised with the bud-burst of its host plants. Due to large individual variation in the length of the pupal period, however, E. autumnata populations exhibit a prolonged period of flight and oviposition. Because the timing of oviposition in autumn is associated with the timing of egg hatch in the following spring, the time window for egg hatch expands and more potential hosts may become attainable. This suggestion was evaluated under field conditions by rearing E. autumnata eggs and larvae on four different hosts.
2. The performance of E. autumnata was measured by using estimates for fecundity (pupal mass) as well as survivorship of eggs and larvae. Based on the availability of foliage and phenological synchrony between larval and leaf development, early-laid eggs and the larvae originating from them were predicted to perform better on the hosts that have early-flushing leaves. On the late-flushing hosts, the larvae that hatched later were predicted to perform better than the larvae that hatched earlier. Half of the trials were exposed to predators and parasitoids, while the rest were conducted inside mesh-bags preventing larval dispersal and mortality due to natural enemies.
3. The results of the experiment did not support the simple predictions. In particular, host-plant quality and natural enemies appeared to operate discordantly between early- and late-laid eggs. Larvae from the late-laid eggs had rapid development during the larval stages and pupated at the same time and with the same pupal mass as the larvae hatched from the early-laid eggs.
4. The results indicate an occurrence of several, unknown selective forces in E. autumnata populations maintaining variation in the length of the pupal period, timing of oviposition, and timing of egg hatch.     

Hatching Response of Meloidogyne incognita acrita to Electric Shock

The influence of electric shock on hatch of Meloidogyne incognita acrita from egg masses taken from roots of ''Acala SJ-I'' cotton (Gossypium hirsutum L.) was studied. Egg masses in tap water were individually placed between the tips of needle electrodes 1 mm apart and exposed to potentials of l, 10, 20, and 60 vdc/mm at 1, 1, 1, and 86 milliamperes dc, respectively, for periods of 2 and 60 seconds. Hatched larvae were counted at five-day intervals for 60 days. Of the eight treatment combinations used, six gave a greater egg hatch than the control. The largest hatch, 520 percent greater than the control, resulted from exposure to 1 vdc/mm for 60 seconds; 60 vdc/mm for 2 and 60 sec decreased egg hatch 11 and 94 percent of the untreated control. Hatched larvae from all treatments except the 60 vdc/mm, 60-second exposure were infective and reproduced on young cotton plants in a glasshouse.     

Field Assessment of Adhesion and Hatch ofChrysoperlaEggs Mechanically Applied in Liquid Carriers

A mechanical technique was evaluated for releasing green lacewing eggs in liquid suspensions. Deposited eggs were enclosed within a circle of nonpoisonous adhesive to protect them from predation and to prevent escape of hatched larvae. Released eggs were monitored daily for 5 days after release by measuring three response variables: adhesion rate of eggs to foliage, hatch rate of eggs, and “yield” of larvae from discharged eggs; “yield” was the product of egg adhesion and egg hatch. Factors tested were: egg conditioning prior to release (incubated or refrigerated), carrier (distilled water or commercial carrier solution), application technique (mechanical or hand application), and row facing (North or South). Release technique did not significantly effect egg hatch on any day. Conditioning eggs prior to release had the greatest effect on hatch of eggs and resulting yield of larvae during the 5-day monitoring period. Carrier had a significant effect on adhesion of eggs to leaves and hatch of eggs. Commercial carrier solution increased egg adhesion but decreased egg hatch compared to water. Overall mean yield of larvae from incubated eggs distributed mechanically was not significantly different for eggs suspended in water (36.4% on day 5 post-release) and for eggs suspended in commercial carrier solution (36.1% on day 5 post-release). Hand-applied eggs had a higher hatch and subsequent yield of larvae than mechanically released eggs; however, the hand technique was labor intensive.     

Interannual differences in growth and hatch date distributions of age-0 year walleye pollock Theragra chalcogramma (Pallas) sampled from the Shumagin Islands region of the Gulf of Alaska, 1985 – 2001

Daily increments of age-0 year walleye pollock Theragra chalcogramma otoliths from 1985 to 2001 were examined for interannual variability in growth and hatch dates. Fish were collected in summer and autumn surveys near the Shumagin Islands in the western Gulf of Alaska (GOA). Hatch date distributions of these fish were compared with hatch dates of larvae from spawning aggregations located in Shelikof Strait and the Shumagin Islands based on their spawning times. The hatch date distributions of age-0 year fish captured from the Shumagin Islands area were similar to those observed for larvae from the Shelikof Strait spawning group. Age-0 year fish whose hatch dates corresponded to the Shumagin Islands spawning, which occurred earlier in January and February, were not found. Sea surface temperature was associated with variability in hatch date distribution and growth.     

In vitro development of the fish parasite Hysterothylacium aduncum from the third larval stage recovered from a host to the third larval stage hatched from the egg

Anisakids are parasitic nematodes of fish worldwide, producing economic and human health concerns. It is thus important to ascertain their in vitro life cycle in laboratory studies. Here we describe the in vitro development of third-stage larvae (L3) of Hysterothylacium aduncum isolated from blue whiting Micromesistius poutassou, to the hatching of L3 from eggs obtained from H. aduncum worms grown in GLIT medium (a modified mixture of Yaeger's LIT [Liver Infusion Tryptose] and Grace's media) at pH 4.0, 13 degrees C and with 5% CO2 in air. Under these conditions, L3 recovered from fish developed to mature adults (3.4 to 6.2 cm in length), with oviposition starting from Day 26 in culture. Fertilized eggs (mean size 64 x 52 microm) had a thick, rugose eggshell and were larger than unfertilized ones (mean size 49 x 42 microm), whose eggshells were thin and smooth. Eggs laid during the first and second week of oviposition, and maintained in 2.8% NaCl solution at 13 degrees C, developed to L3. Under these maintenance conditions, between 20.6 and 52.5% of the eggs laid during the first week developed into larvae. Motile larvae, enclosed in a sheath, hatched from between 2 and 11% of these eggs. The larvae started to hatch 23 d after deposition. These larvae were 144 to 215 microm in length, enclosed in a 237 to 305 microm-long sheath. This GLIT culture medium may help to study the biology of this and other anisakids.     

Modulation of digestive enzyme activities during ontogeny of Labeo rohita larvae fed ascorbic acid enriched zooplankton

The effect of supplementation of ascorbic acid through enriched zooplankton [10%, 20% and 30% ascorbyl palmitate (AP) inclusion in diet of zooplankton] on different digestive enzyme activities during ontogeny of Labeo rohita larvae was studied from 4 day to 15 day post hatch. Ascorbic acid (AA) content in different groups of unenriched (8.6+/-0.71) and enriched zooplankton were, 750+/-29.3, 1409.1+/-45.5, 2009.21+/-199.2 mug/g respectively on dry matter basis with differences (P<0.05) between the treatments. A difference (P<0.05) was found in tissue AA level in different dietary groups. Low amylase, protease, lipase and alkaline phosphatase activities were present in rohu larvae from the mouth opening stage which showed increasing trend with the age of larvae and increasing dietary AA content. A clear dose-dependent modulation of digestive enzyme activities in response to 10%, 20% and 30% AP enriched zooplankton feeding was evidenced from positive correlations between dietary AA content with magnitude of elevation of enzyme activity in different groups. There were 57, 55, 29.2 and 2 fold increases in amylase activity; 7.35, 7.02, 4.43 and 2.73 fold increases in protease activity; 45.636, 41.50, 19.83 and 13.69 fold increases in lipase activity and 6, 5, 3, and 2 fold increases in alkaline phosphatase activity observed in the 15th day post hatch larvae fed 20%, 30%, 10%AP enriched and normal zooplankton respectively, than 4-day post hatch larvae of the respective groups. Enzyme activities were also positively correlated with specific growth rates of wet weight of rohu larvae at the 15th day post hatch. Increased AA might have played an important role in advancing morphological transformation of the digestive tract, protecting gastric mucosa and accelerating growth by the process of tissue formation, which necessitated the requirement of more nutrient thereby, increasing digestive enzyme activity. The regulatory role of AA in the modulation of different digestive enzymes activity and its physiological consequences of nutrient digestibility and utilization during ontogenesis could be extrapolated for better nutrient management of the larvae.     

Necator americanus: temperature, pH, light, and larval development, longevity, and desiccation tolerance

The effect of incubation temperature and pH on the hatch rate of eggs of Necator americanus, and the desiccation tolerance of the resulting infective stage-3 larvae were investigated in the laboratory under controlled conditions. Hatching did not occur below 15 C and above 35 C. A 21% hatch rate was obtained at 15 C while a 10.6% hatch rate was obtained at 35 C. The highest hatch rate (93.7%) was obtained at 30 C. The optimum pH for hatching was 6.0, but the larvae did not reach the infective stage. Incubation temperature of the eggs affected the longevity and desiccation tolerance of resultant infective larvae. Larvae hatched at 30 C and maintained at 26 C under bright fluorescent light had a 50% survival time (S50) of 4 days. In the dark or shade, the S50 for larvae raised at 30 C was 5 weeks, while that of larvae hatched at 20 C was 7 weeks. Incubation temperature also affected the desiccation tolerance of larvae. Larvae developed at 20 C were more resistant to desiccation at various relative humidity values than larvae hatched at 30 C.     

Effects of gamma radiation on spermatogenesis and fertility of male Amblyomma americanum (Acari: Ixodidae)

Amblyomma americanum males were treated with 0.5, 1, 2, 3, 4, 8, and 16 krad of gamma radiation. Testes of ticks treated with 2, 3, 4, 8, and 16 krad were smaller than those of ticks irradiated at lower levels and controls. No recognizable alteration in timing of spermatogenesis was noted among the different radiation groups, but severe breakdown and depletion of germinal cells was noted at 4, 8, and 16 krad. Percent hatch of larvae from crosses of irradiated males and untreated females decreased with increasing radiation level. No hatch was observed from eggs of females mated to males treated at 2 krad or higher.     

Effects of live food and formulated diets on survival, growth and protein content of first-feeding larvae of Plelteobagrus fulvidraco

In recent years, much progress has been made in the rearing of fish larvae fed only artificial diets. A preliminary study was made in an attempt to evaluate the effects of live food and formulated diets on survival, growth and body protein content of first‐feeding larvae of Plelteobagrus fulvidraco. Three test diets varying in protein level were formulated: Feed 1 containing 45% protein, Feed 2 with 50% protein and Feed 3 with 55% protein. Larvae fed live food (newly hatched Artemia, unenriched) were the control. The experiment started 3 days post‐hatch and lasted for 23 days. At the end of the 23‐day trial, survival was best in the control group (65.6%) whereby the final body weight and specific growth rate (SGR) were significantly lower than those in the test feed groups. At the same time, coefficients of variation for SGR and final body weight in the test groups were significantly higher than those in the control. Whole body protein content in all treatments showed a similar tendency during development: significantly higher 3 days post‐hatch, then decreasing significantly, and then increasing unstatistically 10 days post‐hatch. All results suggest that live food is still better for first‐feeding larvae of P. fulvidraco, since live food leads to healthier larvae growth.     

Cryopreservation of common carp (Cyprinus carpio) sperm in 1.2 and 5 ml straws and occurrence of haploids among larvae produced with cryopreserved sperm

Experiments were carried out on the cryopreservation of common carp (Cyprinus carpio) sperm in order to test the suitability of using 1.2 and 5 ml straws and to investigate the ploidy of malformed larvae found among the hatched progeny. In the first set of experiments, the effect of freezing time was investigated on the hatch rate of embryos. The highest hatch rate for 1.2 ml straws was 69+/-16% at the freezing time of 4 min, and 39+/-27% for 5 ml straws at 5 min. In the second set, the effect different egg volumes fertilized with one straw of sperm on the hatch rate and the rate of malformed larvae was investigated. The highest hatch rate with 1.2 ml straws (86+/-12%) was observed when 10 g of eggs were fertilized with one straw, whereas with 5 ml straws the hatch rate was highest (65+/-18%) when 40 g of eggs were fertilized. The highest rate of malformed larvae (15+/-9%) was found in the control, whereas the highest rate of malformed larvae among the groups fertilized with cryopreserved sperm (13+/-7%) was found in the 1x dose group fertilized with 5 ml straw. The chromosome numbers of malformed larvae were investigated and haploids were found among those hatched from eggs fertilized with cryopreserved sperm whereas only diploids were found in the controls.     

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.     

Effects of oximecarbamate, organophosphate and benzimidazole nematicides on life cycle stages of root-knot nematodes, Meloidogyne spp.

A study was made of the effects of concentrations of 2 to 32 ppm of oximecarbamate, organophosphate and benzimidazole nematicides on the hatch, larval viability and migration of Meloidogyne javanica, M. incognita and M. hapla and on development of M. javanica in roots. Aldicarb at less than 8 ppm had little effect on hatch and methomyl markedly affected only the hatch of M. hapla. As little as 2 ppm of fenamiphos or thionazin markedly reduced hatch of all three species but less than 8 ppm ethoprophos significantly reduced only the hatch of M. incognita and phorate had little effect on hatch. Benomyl and thiabendazole had no significant effects on hatch. When egg masses of M. incognita were transferred from nematicides which suppressed hatch to water, hatching occurred, but aldicarb, fenamiphos, ethoprophos and thionazin significantly reduced total hatch. None of the nematicides killed larvae of the three species immersed in 16 and 32 ppm solutions of them for 3 days. Aldicarb at 2 ppm reduced migrations of all three species; the effects of methomyl, fenamiphos or thionazin on migration varied according to species, while phorate, ethoprophos, benomyl or thiabendazole had little or no effect on migration. Aldicarb or thionazin at 2 ppm stopped development of M. javanica in roots of tomato seedlings while methomyl, ethoprophos or fenamiphos at 4 ppm reduced development by 60% and at 8 ppm of ethoprophos or fenamiphos or 16 ppm of methomyl, development was stopped. Phorate had little effect on development and benomyl or thiabendazole had no effect. Nematicide concentrations which reduced development prevented the normal orientation of larvae in the roots and reduced or prevented giant cell formation.     

Induction of dominant lethals with ethyl methane-sulfonate in male germ cells of mulberry silkwork, Bombyx mori l.

Sensitivity of male germ cells in the mulberry silkworm, Bombyx mori L., to ethyl methanesulfonate (EMS) was determined by treating newly emerged 5th- instar larvae, and 2-day- and 7-day-old pupae with 3 concentrations, 0.05, 0.1 and 0.15%, of the mutagen. The frequency of dominant-lethal mutations induced by EMS treatment was used as the parameter for the study. Spermatids and spermatozoa were markedly sensitive to EMS. Statistical analysis confirmed that differences in respect of percentage of egg hatch among the 3 different treatments as well as the interactions between the 3 factors, e.g. stages, hatchability and EMS treatment, were highly significant.     

Biochemical composition during growth and starvation of early larval stages of cultured spiny lobster (Jasus edwardsii) phyllosoma

We examined biochemical changes accompanying feeding and starvation from hatch to Stage VI (day 74 after hatch) in spiny lobster, Jasus edwardsii, phyllosoma larvae. Larval dry weights (dw) increased 17-fold from hatch (80+/-1 microg) to Stage VI (1415+/-44 microg). Larvae starved for 6-11 days at Stages II, IV and VI were 14-40% lighter than their fed counterparts fed enriched Artemia. The increases and losses in total dry weight during feeding and starvation were associated with changes in the content of protein (constituting 31.4-41.7% of dw) and carbohydrate (constituting 2.6-5.3% of dw), while larger changes in lipid content indicated its greater importance as an energy substrate. Lipid content increased from 7.9% of dw at hatch to its highest of 12.5% at Stage IV, but declined by 50% or more during starvation. This suggests that protein, carbohydrate and lipid are all important energy stores, although lipids are catabolized at a greater rate during food deprivation. The principal lipid class was polar lipid (PL; 79-92% of total lipid), followed by sterol (ST; 6-20%), with triacylglycerol and other lipid classes at <2%. PL were catabolized and ST were conserved during starvation. Changes in the fatty acid (FA) profile had mostly occurred before the first moult at day 8 after hatch, with gradual changes thereafter to Stage VI, reflecting their abundance in the Artemia diet. There was some conservation of the major essential FAs, 20:4n-6, 20:5n-3, 22:6n-3, and the FA profile showed large gains in the C(18) polyunsaturated FA, 18:1n-9, 18:2n-6. Ascorbic acid content increased 10-fold from hatch to the end of Stage I (36 and 333 microgg(-1) dw, respectively), while the content at the end of Stage II was higher in fed than that in starved larvae (439 and 174 microgg(-1) dw, respectively). Our study will assist in the development of alternatives to nutritionally incomplete diets, such as live ongrown Artemia, to meet the requirements of phyllosoma in culture.     

The community ecology of Aedes egg hatching: implications for a mosquito invasion

Abstract. 1. A recently introduced treehole mosquito from Asia, Aedes albopictus , is spreading throughout eastern North America, especially in tyre-refuse piles. Previous studies have identified inhibitory effects of larvae on egg hatch as a potential population regulatory mechanism within Aedes. Larva-egg interactions may also occur between species. This experiment assesses the ability of larvae of A. albopictus and two possible competitors in North America, A.triseriatus and A.aegypti , to suppress hatching of conspecific and congeneric eggs.
2. We exposed eggs of each species to varying combinations of larval species and density for 24h and assessed subsequent hatch rates. Aedes albopictus eggs exhibited the lowest level of inhibition when exposed to high larval densities; moreover, at the lowest larval density they imposed the most intense interspecific hatch inhibition.
3. Discretionary hatching in response to larval density may influence community composition by promoting the spread of A.albopictus , perhaps even leading to its dominance within North American Aedes communities.     

Direct and indirect effects of simulated calcareous dredge material on eggs and larvae of pink snapper Pagrus auratus

The direct and indirect effects of a simulated, calcarenite‐based dredge material on eggs and larvae of pink snapper Pagrus auratus were assessed. Direct effects were assessed by measuring hatch rate or survival of eggs and pre‐feeding larvae, respectively, over a range of concentrations and exposure durations. Exposure of eggs to suspended solid concentrations up to 10 000 mg l^?1 for 24 h did not affect egg buoyancy or hatch rate, despite sediment adherence occurring at the two highest concentrations tested. Newly hatched larvae, whose mouths were still closed, were relatively tolerant of suspended solids, with a 12 h lethal concentration resulting in 50% mortality, LC₅₀, of 2020 mg l^?1 and a first observable effect concentration of 150 mg l^?1. Once the larvae's mouths opened, tolerance was significantly reduced, with a 12 h LC₅₀ of 157 mg l^?1 and a first observable effect concentration of 4 mg l^?1. Tolerance of larvae to suspended solids was negatively correlated with suspended solids concentration and exposure time, with exposure durations of ≤6 h being significantly less detrimental than those of 9 h or more. Indirect effects to larvae were assessed by measuring ingestion of copepod nauplii by 10 and 15 days post‐hatch (dph) larvae at sediment concentrations from 0 to 200 mg l^?1 in 50 mg l^?1 increments over 4 h. Ingestion was not significantly affected by sediment for 10 dph larvae, but by 15 dph, sediment had a far greater impact on ingestion, with larvae in all sediment treatments eating significantly fewer copepods than those in the control.     

Egg hatching of mosquitoes Aedes caspius and Ae. vittatus stimulated by water vibrations

Simple laboratory experiments with eggs of wild-caught Aedes caspius (Pallas) and Aedes vittatus (Bigot) mosquitoes (Diptera: Culicidae) demonstrated that egg hatch rates (i.e. percentage hatching daily) increased significantly in response to water vibrations caused by finger-tip drumming on containers of submerged eggs. When disturbed by daily vibrations for only 30 s, eggs hatched sooner than when they remained flooded or were periodically drained and reflooded without agitation. For autogenous Ae. caspius from saltmarsh, egg hatch rates were three- to four-fold greater for groups of 50 eggs than for solitary eggs, possibly due to chemicals (functional kairomones) emanating from other eggs or larvae. For anautogenous Ae. vittatus from freshwater rock-pools, tapping the container daily (for 30 s each 2 h in 8 h) hatched 93% of eggs, compared with only 42% hatch of eggs agitated only when first flooded or 55% hatch of eggs dried and reflooded three times during 17 days without other disturbance. It is concluded that, apart from flooding and other factors, vibrations - simulating the patter of rainfall provide a very significant hatching stimulus for Aedes eggs.