Biological effect of low doses of alkylating agents in drosophila studies

The low dose (0.05-0.1 mM) influence of alkylating agents on germ cell survival and male fertility, the level of embryonic and postembryonic lethality as well as the sex-linked recessive lethal (SLRL) frequency induced by high alkylating agent doses was studied in Drosophila melanogaster. The pretreatment of adult males with low doses of methyl and ethyl methanesulfonate (MMS and EMS) did not change or even enhanced EMS cytotoxicity and mutagenicity in both mature sperm and premeiotic cells. On the contrary, the low EMS dose pretreatment of larvae protected them against higher mutagen doses increasing male fertility, decreasing embryonic and postembryonic lethality in F1, and leading to three-fold reduction in the SLRL frequency in F2. The adaptive response was dependent on the Drosophila developmental stage exposed to challenge mutagen doses, since the protection was maximal in larvae and practically absent when the high dose was administered to adult males. The adaptive response observed does not seem to be associated with DNA repair, but it is rather due to other protective mechanisms.     

Adaptive response to alkylating agents in the Drosophila sex-linked recessive lethal assay

The adaptive response to alkylating agents was studied in Drosophila assays under various treatment procedures. Pre-treatment of males as well as treatment of females with low doses of EMS (0.05-0.1 mM) did not affect sex-linked recessive lethal (SLRL) rates induced by high doses of this mutagen (10 mM, various feeding duration) in mature sperm cells. Pre-treatment of males with a low dose of MMS (0.1 mM) enhanced mutagenesis induced by the high dose of EMS (10 mM) at different stages of spermatogenesis, the observed effects exceeding the additive action of both mutagens. On the contrary, larval pre-treatment with the adaptive dose of EMS (0.05 mM) resulted in resistance of their germ cells to higher doses of EMS (1 mM). Specifically, offspring production increased while dominant lethality in F(1) as well SLRL frequency in F(2) was significantly reduced as compared with the effects of larval exposure to the challenge dose. Under the conditions tested, the adaptive response of germ cells to alkylating agents was demonstrated in larvae, but not in adult flies.     

Cadmium uptake by rainbow trout, Salmo gairdneri eggs and alevins

The uptake of cadmium by eggs and alevins of rainbow trout from water concentrations of between 0.01 and 50.0 mg Cd 1⁻¹ was investigated. The cadmium content of eggs and alevins increased with time and with exposure concentration. Lower cadmium levels were detected in alevins than in eggs. Most of the cadmium (98%) in the eggs was found to be associated with the egg membrane or chorion. This explains the considerable reduction in cadmium concentration observed in alevins after hatching. Alevins hatching from eggs which had been exposed to cadmium survived longer in cadmium than alevins not exposed as eggs. This suggests that the pretreatment of eggs with cadmium serves some protective function. Behavioural and pathological signs of cadmium poisoning such as erratic swimming and blood clotting in alevins were observed.     

Adult diet and male-female contact effects on female reproductive potential in Mexican fruit fly (Anastrepha ludens Loew) (Diptera tephritidae)

Wild strains of fruit flies (Diptera: Tephritidae) placed into laboratory rearing conditions are subjected to selection pressures caused by the diet, cages, density of flies, and other factors. Selection that changes mating behavior of the strain may result in less effective males released in sterile insect programs. Tests were performed to examine the effects of protein in diet and adult interactions on egg production and mating during sexual maturation of the Mexican fruit fly (Anastrepha ludens Loew) in laboratory cages. Flies were offspring of wild flies collected from Chiapas or Nuevo Leon, Mexico, and reared on Valencia oranges. Experiments demonstrated effects of yeast hydrolysate protein in adult diet and pairing with males on production of mature and immature eggs, numbers of females producing eggs, and mating with females aged 15 d. Addition of protein to 4% fructose in the adult diet approximately tripled mature egg production in females maintained for the total maturation period with an equal number of males. Females that matured without males produced approximately 33% more-mature eggs when fed protein than those fed no protein. Total egg production of females matured without males and fed sugar only or sugar with protein was more than twice that of females matured with males. Tests to examine the effects of male and female diet separately on female egg production showed slightly higher egg production in females fed protein, or females paired with males fed protein, but these differences were not significant. The most definitive effects were that combining wild strain females and males in cages during maturation reduced egg production. This effect was greatest when flies were not fed protein.     

Reduction in egg hatch after a sublethal dose of chlorfluazuron to larvae of the common cutworm, Spodoptera litura

Abstract.  The effects of a sublethal dose of chlorfluazuron on egg hatching in Spodoptera litura are examined under laboratory conditions. When LD₁₀ of chlorfluazuron is topically applied to newly moulted fifth-instar larvae of either sex, a significant reduction in both the number of eggs laid and subsequent hatching is observed after mating but no significant differences in daily of oviposition are observed when compared with the controls. In addition, examination of the unhatched eggs reveals that the number of unfertilized eggs is greater than those that were fertilized but there are significantly more unfertilized eggs laid by treated insects. Interference by chlorfluazuron, transferred by copulation through sperm fluids or ova, appears improbable. All the fertilized unhatched eggs in the treated crosses die at an earlier stage than those of the controls. In the female-treated crosses, the egg size is significantly reduced compared with the control or male-treated crosses. During mating, the treated-males transferred significantly lower-weight spermatophores into the females. The weight of spermatophores transferred by untreated males is the same to both treated and untreated females. The duration of mating is not affected by chlorfluazuron treatment.     

Chromosomal analysis in mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) and methyl and ethyl methanesulfonate (MMS and EMS)

Chromosome aberrations were analyzed at the first-cleavage metaphase of mouse eggs fertilized in vitro with sperm exposed to ultraviolet light (UV) as well as to methyl and ethyl methanesulfonate (MMS and EMS). The frequencies of chromosome aberrations markedly increased with dose of UV as well as with concentration of MMS and EMS. In the UV-irradiation group, the frequency of chromosome-type aberrations was much higher than that of chromatid-type aberrations. About 90% of chromosome aberrations observed in the eggs following MMS and EMS treatment to sperm were chromosome type in which the frequency of chromosome fragments was the highest. The effects of UV on the induction of chromosome aberrations were clearly potentiated by post-treatment incubation of fertilized eggs in the presence of Ara-C or caffeine, but the effects of MMS and EMS were not pronounced by post-treatment of Ara-C or caffeine. The results indicate a possibility that UV damage induced in mouse sperm DNA is reparable in the eggs during the period between the entry of sperm into the egg cytoplasm and the first-cleavage metaphase.     

Inactivation of Salmonella enterica serovar enteritidis in shell eggs by sequential application of heat and ozone

Aims:  To assess the contribution of ozone to lethality of Salmonella enterica serovar Enteritidis in experimentally inoculated whole shell eggs that are sequentially treated with heat and gaseous ozone in pilot-scale equipment.
Methods and Results:  Whole shell eggs were inoculated with small populations of Salmonella Enteritidis (8·5 × 10⁴–2·4 × 10⁵ CFU per egg) near the egg vitelline membrane. Eggs were subjected to immersion heating (57°C for 21 min), ozone treatment (vacuum at 67·5 kPa, followed by ozonation at a maximum concentration of approx. 140 g ozone m⁻³ and 184–198 kPa for 40 min) or a combination of both treatments. Survivors were detected after an enrichment process or enumerated using modified most probable number technique. Ozone, heat and combination treatments inactivated 0·11, 3·1 and 4·2 log Salmonella Enteritidis per egg, respectively.
Conclusions:  Sequential application of heat and gaseous ozone was significantly more effective than either heat or ozone alone. The demonstrated synergy between these treatment steps should produce safer shell eggs than the heat treatment alone.
Significance and Impact of the Study:  Shell eggs are the most common vehicle for human infection by Salmonella Enteritidis. Many cases of egg-related salmonellosis are reported annually despite efforts to reduce contamination, including thermal pasteurization of shell eggs and egg products. Treatment with ozone-based combination should produce shell eggs safer than those treated with heat alone.     

Effect of mating status on the fecundity of a cricket,Teleogryllus emma

Abstract Fecundity in some insects is affected by mating status. The effect of mating status on the fecundity and total egg production of Teleogryllus emma (Ohmachi et Matsumura) (Orthoptera: Gryllidae) was examined in this study. The results showed that the pre‐oviposition period was shorter for amphigonic females than that for virgin females. However, no significant difference in pre‐oviposition was found between amphigonic females and those that had mated with a male with either the phallodeum or testes extirpated. There is no difference in adult longevity between the above four groups. The fecundity and total egg production were much higher in amphigonic females than in those controlled under the three non‐amphigonic treatments. The females of T. emma that mated with the testes‐extirpated males produced more eggs (up to two‐fold more) than both the virgin females and those that mated with the phallodeum‐extirpated males, but there was no difference between them. The fecundity‐enhancing substances transferred from male to female can stimulate the female to produce more eggs, but this stimulation has to occur in collaboration with sperm.     

Fatty acid signatures of female Brünnich's guillemots Uria lomvia suggests reliance on local prey for replacement egg production

The laying of smaller replacement eggs has been described as a time-saving adaptation because chicks generally grow faster once hatched than inside the egg. However, chicks hatched from smaller eggs have, potentially, lower survival. Consequently, the smaller replacement egg represents a benefit to the female in terms of preserving its own condition at a cost to their offspring. We test these ideas by measuring adult mass changes and plasma lipid concentration changes in male and female Brünnich's guillemots Uria lomvia breeding on Coats Island, Nunavut. Though males lost more mass than females, these differences were not significant. Between laying the first and replacement egg, plasma fatty acid concentrations declined in females and increased in males, suggesting that females mobilise less lipid to preserve their condition after laying the replacement egg. In females, plasma lipid concentrations of the dominant fatty acids found in the eggs (16:0 and 18:1) declined between the laying of first and replacement eggs while plasma concentrations of 20:1 increased in both males and females. We compared the fatty acid signatures of first and replacement egg to look for evidence of differences between the lipid sources for their production. Principal component and discriminant function analyses showed that the fatty acid signatures of replacement eggs were closer to the signatures of the local prey than those of first eggs. We suggest that females rely on local sources of energy to a greater degree for the production of the replacement egg than the first egg, but that endogenous reserves of certain nutrients are important for the production of both eggs.     

Long-term effects of methoxyfenozide on the adult reproductive processes and longevity of Spodoptera exigua (Lepidoptera: Noctuidae)

The long-term effects of methoxyfenozide on the longevity and reproductive processes of beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae), adults were assessed after exposure by ingestion. Methoxyfenozide significantly reduced adult male longevity compared with females by 1.1 and 1.5 d at 75 and 150 mg (AI)/liter, respectively. Fecundity decreased by >60% with both concentrations at 72 and 96 h after treatment, but at 48 h, no significant effect was observed. The carbohydrate, protein, and lipid content in the eggs were determined as representatives of the biochemical effects of methoxyfenozide associated with the disruption of reproductive processes. The content of carbohydrates in the eggs laid 48 h at treatment was similar to that of controls, but it increased by approximately 1.5 and 2-fold in eggs laid after 72 and 96 h, respectively, compared with controls (15 microg per egg). Protein content was reduced approximately 2.5 and approximately 3-fold for each treatment concentration, respectively, compared with the controls (25 and 23 microg per egg for 75 and 150 mg [AI]/liter, respectively) in eggs collected 72 and 96 h after treatment. Lipid content significantly decreased by approximately 1.6-fold in both treatment concentrations in eggs collected at 48 and 96 h after treatment compared with the controls (24 and 21 microg per egg for 48 and 96, respectively), but it was similar to controls (approximately 19 microg per egg) at 72 h (approximately 15 microg per egg) for both concentrations. The biochemical effects of methoxyfenozide on S. exigua egg formation detected in this work are consistent with the reduction in fertility observed, as reported previously.     

Ovicidal and adulticidal effects of monoterpenoids against permethrin-resistant human head lice, Pediculus humanus capitis

Abstract.  The improper use of pediculicides containing permethrin has led to the development of resistance. Thus, new alternatives for control are needed. Plant-derived insecticides are attractive alternatives to common chemical insecticides because most of them are environmentally friendly and non-toxic to mammals. The toxic activity of 23 monoterpenoids belonging to several chemical classes was tested against the eggs of permethrin-resistant head lice, Pediculus humanus capitis De Geer (Phthiraptera: Pediculidae). Significant differences in ovicidal action were observed among the tested substances. The most effective chemicals were hydrocarbons and ethers, followed by ketones, alcohols, phenols and esters. A linear relationship between egg mortality and knockdown time (KT₅₀) on adults by the tested components revealed that most of the components were effective on both egg and adult stages. The monoterpenoids described herein are good candidates as effective pediculicides.     

Comparison of dominant lethal and heritable translocation methodologies

Groups of male Alderly Park mice of proven fertility were dosed by gavage for 5 consecutive days per week for 8 weeks or 5 consecutive days only with 100 or 150 mg/kg bodyweight ethyl methanesulphonate (EMS) or by intraperitoneal injection once a week for 8 weeks or once only with 500 mg/kg shikimic acid. Animals dosed in this manner were compared in the dominant lethal and heritable translocation assays. Animals were mated for 2 consecutive weeks following the 8-weak treatment and for 8 consecutive weeks after the 1-week treatment: regimes which were thus non-specific and specific respectively for the stages of spermatogenesis. An additional method of measuring dominant lethality involving counting uterine scars after weaning (Soares (1972) Mutation Res., 16, 425–427) was used and also compared with the conventional method.EMS was clearly confirmed as a mutagen but this was not the case for shikimic acid. For screening purposes the dominant lethal 8-week mating assay was much more efficient in return for the same effort for detecting mutagenic responses than an 8-week mating heritable translocation assay, since the induction of dominant lethal effects paralleled the induction of heritable translocations. 8-week treatment with EMS showed increased dominant lethality but severely reduced fertility and the small numbers of male offspring born made potential heritable effects difficult to assess. The 1-week treatment with EMS produced both dominant lethal and heritable effects. Soares' method can be useful for determining determinant lethal effects in a heritable translocation assay. The “sieving” method of mating to determine partial and total sterility questions the necessity for a negative control in a heritable translocation study.     

Synchronization of larval emergence in winter moth (Operophtera brumata L.) and budburst in pedunculate oak (Quercus robur L.) under simulated climate change

1. The hypothesis that a 3 °C elevation in temperature and doubled CO₂ concentration would have no effect on the synchronization of winter moth egg hatch with budburst in oak was tested by comparing the separate and interactive effects of ambient and elevated (+ 3 °C) temperature and ambient and elevated (doubled to 340 p.p.m.) CO₂ in eight experimental Solardomes. In addition, an outdoor control was compared with the ambient temperature/CO₂ treatment combination.
2. Elevated temperature accelerated darkening (preceding egg hatch by about 5–10 days) and hatching of eggs developing off the trees; elevated CO₂ had no effect. The same effects were observed in eggs developing on the trees.
3. Within treatments, date of egg hatch was the same on trees with early or late budburst.
4. Egg darkening and budburst were closely synchronized at both ambient and elevated temperatures.
5. Both eggs and trees required fewer cumulative heat units (day degrees > 4 °C), for hatching and budburst, respectively, at ambient than elevated temperatures. The requirements in the outdoor control treatment were similar to those in the ambient Solardome treatment.
6. Egg hatch between 10 and 25 °C, on a temperature gradient in the laboratory, required a constant number of heat units; fewer were required below 10 °C.
7. Elevated temperatures, in the Solardomes and the field, delayed adult emergence from the pupae.
8. The results suggest that a general increase in temperature with climatic change would not affect the closeness of the synchronization between egg hatch of winter moth and budburst of oak.     

Cannibalism and protective behavior of eggs in Arctic charr (Salvelinus alpinus)

From video recordings of spawning events, we quantified protective and cannibalistic behavior of Arctic charr occurring immediately after spawning. The number of fish cannibalizing on stray eggs was examined regarding (a) whether more than one male shed milt during the spawning event, that is, whether sperm competition occurred, (b) whether the sperm competition included few or many males, that is, the intensity of sperm competition, and (c) the density of fish at the spawning site. Response behavior toward egg cannibalism was also examined among females and dominant males in order to determine any parental investment toward protecting the eggs after spawning. Cannibalistic behavior was seen in almost 50% of the spawnings, and the multiple spawning events showed the highest numbers of fish cannibalizing on eggs. Both the number of males releasing milt and the number of fish approaching the spawning site were positively correlated with egg cannibalism. Sperm competition was, however, not a prerequisite for egg cannibalism. Although we also observed partial filial cannibalism, protective behavior of eggs was seen both among dominant males and females, suggesting that charr actually conduct parental care.     

Effect of relative humidity on development, survival and reproduction of the predatory lacewing Dichochrysa prasina (Neuroptera: Chrysopidae)

The effect of five constant relative humidity (RH) regimes (12%, 33%, 55%, 75% and 94%) on the preimaginal development and adult longevity and reproduction of Dichochrysa prasina Burmeister was studied at 26 ± 1 °C and a photoperiod of 16:8 (L:D) under laboratory conditions. For the experiments, larvae were fed individually on eggs of the flour moth Ephestia kuehniella (Zeller), whereas adults on a liquid diet consisting of water, yeast hydrolysate, sugar and honey. The experimental insects were maintained individually in sealed plastic containers with saturated water salt solutions at the base to control RH. Preimaginal developmental time from egg to adult for females and males was significantly shorter at 75% and 94% RHs than at other RHs in the range from 12% to 55%. Percentages of adult emergence were similar at all RHs tested and varied from 52.5% to 75.0%. Adult females that had no access to water and were maintained at 12%–75% RH lived on average less than two weeks and laid no eggs, whereas at 94% RH they lived longer and each laid on average 201 eggs. When the females had periodic access to a water source a significant increase in both longevity and egg production occurred and the estimated intrinsic rate of increase was high, irrespective of RH. These results could be useful for the mass-rearing of D. prasina and for better understanding its population dynamics under field conditions.     

Variation in behavioural response to oxygen stress by egg-tending males of parapatric fluvial and lacustrine populations of a landlocked goby

The behavioural response of egg-tending males from parapatric fluvial and lacustrine populations of a landlocked goby Rhinogobius sp. (the orange form) to oxygen stress was compared in laboratory experiments. The natural spawning locations of these populations (rapids of tributary rivers and lakeshore of Lake Biwa, respectively) differ in dissolved oxygen concentration and its variability. Males of both populations spent a longer period of time in fanning behaviour under low dissolved oxygen conditions (4·5–5·0 mg l⁻¹), where >90% of eggs without paternal care died before reaching eyed stage, relative to fully saturated dissolved oxygen conditions (8·0–8·5 mg l⁻¹). Lacustrine males, who occasionally encounter oxygen stress (<2 mg l⁻¹) in their natural habitat, fanned eggs for a longer time period than fluvial males. The time difference in fanning behaviour between the two oxygen conditions was greater for lacustrine than fluvial males. Survival rate in the lower oxygen condition was higher for eggs tended by lacustrine males than those tended by fluvial males, probably due to this difference in fanning activity. These results showed that the response to oxygen stress differs between the two populations and, moreover, as both populations behaved adaptively in responding to the reduction in dissolved oxygen, contiguous habitats may have distinct natural selective pressures. It is suggested that regulation of egg fanning activity is strongly favoured by natural selection in unpredictable environments.     

Effects of egg age of Spartocera dentiventris (Berg) (Hemiptera: Coreidae) on parasitism by Gryon gallardoi (Brethes) (Hymenoptera: Scelionidae)

To evaluate the effect of egg age of Spartocera dentiventris (Berg) on the parasitism by Gryon gallardoi (Brethes), groups of 12 eggs of 2, 3, 4, 5, 6, 7, 8, and 12 days old were exposed to one female of G. gallardoi on a test tube, with the wasp observed for 2h subsequently. Later, the eggs were individualised and incubated (25 +/- 1 degrees C; 12h photoperiod) until adult emergence. To identify the preferred age, groups of 12 eggs of 1, 3, 5, 7 and 12 days of age were simultaneously exposed to a parasitoid female on an arena for 2h. These groups were also individualised and incubated until parasitism was confirmed. On all ages tested parasitism was at about 88.5 +/- 2.64%, with a reduction in adult parasitoid emergence with age increasing (P < 0.01). The development time increased with increasing egg age (P < 0.01), with males developing faster. Parasitoid sex ratio for all ages tested was female-biased (0.49:1), with the proportion of males invariant with host age (P > 0.05). When the egg groups were exposed simultaneously, the female preferred eggs one and three days old. The results indicate a preference of G gallardoi for younger eggs of S. dentiventris, with host egg age exerting a negative effect on development time and offspring emergence of the parasitoid.     

Resurrection of Aedes (Halaedes) ashworthi Edwards 1921: Morphological Characterisation and Separation from its Sibling Aedes (Halaedes) australis (Erichson) 1842 (Diptera: Culicidae)

The result of population crosses of members of the Ae. australis complex and the examination of the male genitalia of these populations have shown that Ae. ashworthi Edwards 1921 should be reinstated as a valid species. the parent females laid viable eggs when cross mated to males of different species, but the resulting F₁ females and males were largely sterile. When F₁ females were blood-fed and examined 3–8 d later, usually fewer than 10 ovarioles per ovary developed recognisable follicles and none of these formed eggs. F₁ males in nearly all cases possessed half-size testes, and there was no spermatid development in the reduced testes. A few males had normal-sized testes, but the spermatids were mostly tailless. the male of Ae. ashworthi can be separated from Ae. australis by the shape of the gonocoxite and the length of the gonostylus of the genitalia. the larvae of both species are identical, and the females differ only in the colour of some scale patches and the integument, which may prove to be too variable to be useful when more populations are compared. the morphological similarity of the two mosquitoes as larvae and adults supports the proposal that they should be recognised as sibling species, with Ae. ashworthi as the derived member of the Ae. australis complex. It is clear from morphological evidence, the viability of the F₁ hybrids and the unusual habitat occupied by both species, that they have diverged recently.     

Sugar transport in leaf discs of Phaseolus coccinius

Kinetic profiles for sucrose, glucose and 3-OMG glucose were determined in leaf discs of Phaseolus coccinius L. (cv. Scarlet). All three sugars exhibited identical uptake kinetics. At sugar concentrations below 25 m M , transport was due to an active, carrier-mediated transport system. A linear component was the dominant mode of uptake at sugar concentrations above 25 m M . Sucrose and glucose carriers were specific for these sugars, since no uptake inhibition was observed from competing sugars. Sucrose was not hydrolyzed by leaf tissue because the label in asymetrically labeled sucrose was not randomized. Furthermore, no label was present in hexose fractions when tissue was incubated with [⁸⁴C]-sucrose. Therefore, [¹⁴C]-sucrose uptake did not reflect hexose uptake.
Both saturable and linear components of uptake contribute significantly to total uptake rates. The former, however, is more important when apoplastic sugar concentrations are low. The molecular nature of the linear component is not well understood but accounts for most of the uptake at high sugar concentrations.     

Spermatophore formation and regulation of egg maturation and oviposition in female Heliothis virescens by the male

This study was undertaken, using various surgical manipulations, to examine the role and functions of accessory sex glands and testes in Heliothis virescens in spermatophore formation and stimulation of egg maturation in mated females. Normal females mated to accessory sex glandectomized (-ASG) males produced fewer eggs and retained most of their eggs in ovarioles compared with females mated to sham-operated (+ASG) or normal males. The ASG are the source of the components for spermatophore formation in H. virescens. Females mated to castrated (-Testes) males showed similar pattern of egg production as did females mated to -ASG males. Females mated to -Testes males and those mated to sham-operated (+Testes) or normal males were found to have spermatophore in the bursa after uncoupling. Normal females mated to allatectomized (-CA) males developed similar numbers of total eggs as did females mated to sham-operated (+CA) males and normal males.