UV reflectance of eggs of brown-headed cowbirds (Molothrus ater) and accepter and rejecter hosts

Many hosts of obligate brood parasites accept parasitic eggs despite the high costs of parasitism. Acceptance is particularly perplexing in brown-headed cowbird (Molothrus ater) (hereafter “cowbird”) hosts because the eggs of cowbirds and most hosts do not appear to match closely in visual characteristics detectable by humans. However, recent evidence suggests that parasite and host eggs may match in their ultraviolet (UV) reflectance, undetectable by humans, and that birds may use UV signals for egg discrimination. We determined whether egg colour matching in UV reflectance separates accepters and rejecters of cowbird parasitism by comparing the total UV (300–400 nm) reflectance of the eggs of 11 host species to cowbird eggs. Eggs of three of five accepter species and five of five rejecter species differed significantly from cowbird eggs in UV reflectance. We found no significant difference in the UV reflectance of the eggs of three closely related pairs of accepter and rejecter species. There also was no significant difference in the UV reflectance of cowbird eggs laid in nests of five host species, and the UV reflectance of cowbird eggs was not significantly correlated with that of host clutches. Thus, we found no support for the UV-matching hypothesis in brown-headed cowbirds and UV reflectance does not appear to separate accepters and rejecters of parasitism. Differences in UV reflectance between cowbird and host eggs, however, provide potential cues for use in egg discrimination. Experimental testing is needed to determine the relative importance of UV reflectance compared to other visual cues.     

Two UV-sensitive targets in dorsoanterior specification of frog embryos

Previous work has shown that ultraviolet (UV) irradiation of fertilized frog eggs yields embryos that lack dorsal and anterior structures. The eggs fail to undergo the cortical/cytoplasmic rotation that specifies dorsoventral polarity, and they lack an array of parallel microtubules associated with the rotation. These eggs can be rescued by tilting with respect to gravity, and normal dorsoanterior development occurs. We find here that UV irradiation of Xenopus prophase I oocytes or Rana metaphase I oocytes also causes the dorsoanterior deficient syndrome, but the UV target is different from that in fertilized eggs. Tilting eggs, irradiated as oocytes, with respect to gravity, does not rescue dorsoanterior development, although lithium treatment does. The UV dose required to produce dorsoanterior deficiency for Rana metaphase I oocytes is much less than that for fertilized eggs, and the oocytes can form the array of parallel microtubules and undergo the cortical/cytoplasmic rotation after fertilization. Despite these features of normal development, no dorsoanterior structures form. While the UV target in fertilized eggs is thought to be the parallel microtubules (Elinson & Rowning, 1988; Devl Biol. 128, 185-197), the UV target in the oocytes may be a dorsal determinant.     

Inactivation of single-celled Ascaris suum eggs by low-pressure UV radiation

Intact and decorticated single-celled Ascaris suum eggs were exposed to UV radiation from low-pressure, germicidal lamps at fluences (doses) ranging from 0 to 8,000 J/m2 for intact eggs and from 0 to 500 J/m2 for decorticated eggs. With a UV fluence of 500 J/m2, 0.44-+/-0.20-log inactivation (mean+/-95% confidence interval) (63.7%) of intact eggs was observed, while a fluence of 4,000 J/m2 resulted in 2.23-+/-0.49-log inactivation (99.4%). (The maximum quantifiable inactivation was 2.5 log units.) Thus, according to the methods used here, Ascaris eggs are the most UV-resistant water-related pathogen identified to date. For the range of fluences recommended for disinfecting drinking water and wastewater (200 to 2,000 J/m2), from 0- to 1.5-log inactivation can be expected, although at typical fluences (less than 1,000 J/m2), the inactivation may be less than 1 log. When the eggs were decorticated (the outer egg shell layers were removed with sodium hypochlorite, leaving only the lipoprotein ascaroside layer) before exposure to UV, 1.80-+/-0.32-log reduction (98.4%) was achieved with a fluence of 500 J/m2, suggesting that the outer eggshell layers protected A. suum eggs from inactivation by UV radiation. This protection may have been due to UV absorption by proteins in the outer layers of the 3- to 4-microm-thick eggshell. Stirring alone (without UV exposure) also inactivated some of the Ascaris eggs (approximately 20% after 75 min), which complicated determination of the inactivation caused by UV radiation alone.     

Dark nests and egg colour in birds: a possible functional role of ultraviolet reflectance in egg detectability

Owing to the conspicuousness of ultraviolet (UV) colour in dark environments, natural selection might have selected UV egg coloration because it would enhance egg detectability by parents in murky nests. Here, we tested this hypothesis by using comparative and experimental approaches. First, we studied variation in egg coloration of 98 species of European passerines measured using UV-visible reflectance spectrometry (300-700nm) in relation to nesting habits. Analyses based on raw data and controlling for phylogenetic distances both at the species and the family levels revealed that hole-nester species produced eggs with higher UV reflectance than those nesting in open habitats. The experimental approach consisted of the manipulation of UV reflectance of the experimental eggs introduced outside the nest-cup of the hole-nester spotless starling Sturnus unicolor and the study of the retrieval of these eggs. Ultraviolet-reflecting eggs (controls) were more frequently retrieved to the nest-cup than non-reflecting (-UV) eggs. These results were not due to '-UV' eggs being recognized by starlings as parasitic because when a parasitic egg is detected, starlings removed it from the nest-box. Therefore, these results are consistent with the hypothesis that UV egg colours are designed to provide highly detectable targets for parent birds in dark nest environments.     

The effect of a commercial UV disinfection system on the bacterial load of shell eggs

AIMS: To study the effect of UV irradiation on the bacterial load of shell eggs and of a roller conveyor belt. METHODS AND RESULTS: The natural bacterial load on the eggshell of clean eggs was significantly reduced by a standard UV treatment of 4.7 s; from 4.47 to 3.57 log CFU per eggshell. For very dirty eggs no significant reduction was observed. Eggs inoculated with Escherichia coli and Staphylococcus aureus (4.74 and 4.64 log CFU per eggshell respectively) passed the conveyor belt and were exposed to UV for 4.7 and 18.8 s. The reduction of both inoculated bacteria on the eggshell was comparable and significant for both exposure times (3 and 4 log CFU per eggshell). Escherichia coli was reduced but still detectable on the conveyor rollers. The internal bacterial contamination of eggs filled up with diluent containing E. coli or S. aureus was not influenced by UV irradiation. Conclusions: There is a significant lethal effect of UV irradiation on the bacterial contamination of clean eggshells and recent shell contamination, contamination of rollers can be controlled and the internal contamination of eggs is not reduced. SIGNIFICANCE AND IMPACT OF THE STUDY: The penetration of UV into organic material appears to be poor and UV disinfection can be used as an alternative for egg washing.     

UV reflectance as a cue in egg discrimination in two Prinia species exploited differently by brood parasites in Taiwan

Birds are capable of seeing the ultraviolet light (UV) spectrum and as a consequence have evolved UV‐reflective structures with signalling functions. Avian eggs also reflect in the UV spectrum but the importance of UV egg matching in egg rejection decisions has been equivocal. Here we conducted egg rejection experiments in the congeneric and sympatrically breeding Yellow‐bellied Prinia Prinia flaviventris and Plain Prinia Prinia inornata in Taiwan to assess the role of UV as a cue in egg discrimination. Yellow‐bellied Prinia is a host of Oriental Cuckoo Cuculus optatus, whereas Plain Prinia is not. We coated one prinia egg in the experimental clutches with a cream containing a UV‐blocking agent, while the rest of the eggs were coated with cream only. We also experimentally parasitized prinias with non‐mimetic model eggs with reduced UV reflectance. Yellow‐bellied Prinia and Plain Prinia rejected their own UV‐blocked eggs in 18.2 and 8.3% of the experiments, respectively, and the difference was not significant. However, Yellow‐bellied Prinia rejected 100% of the non‐mimetic eggs, whereas the Plain Prinia rejected only 5%. Hence, UV reflectance alone is a cue in egg discrimination, but the importance of reflectance outside the UV spectrum in these two prinia species is much more responsive to selection as a consequence of brood parasitism.     

Oviposition diapause and other factors affecting the egg-laying of Phlebotomus papatasi in the laboratory

Stimuli which modulate oviposition of P. papatasi were investigated to improve insectary breeding efficiency. Oviposition and survival of gravid females were observed weekly during April-December 1987, in plastic cages at 28 +/- 1 degrees C with L:D 17:7. Oviposition of controls was subject to seasonal variation despite the relatively uniform insectary conditions. From April to mid-October (summer), mean weekly oviposition ranged from 11.6 to 18.6 eggs per fly, dropping to 1.4 eggs/fly in November (winter). Monthly yields of eggs were found to correlate with the seasonal cycle of ultraviolet (UV) radiation in sunlight. This was attributed to an endogenous rhythm since the flies were not normally exposed to UV under insectary conditions. Short exposure to UV sources of 254 nm and 312 nm, but not white light, raised the low rate of oviposition in October-November from 1.4 to 16.8-29.6 eggs/female/week, but female mortality also increased highly significantly. It is suggested that the seasonal oviposition cycle of P.papatasi is set by levels of UV irradiation. In the warm season oviposition was promoted, as compared to controls, by furrows in plaster of Paris lining the bottom of cages (29.6 v. 10.2 eggs/female) and by cow manure in the cages (39.7 v. 18.2 eggs/female), but the combination of both stimuli gave no greater fecundity (40.9 v. 20.9 eggs/female). Oviposition decreased when larvae were present (3.9 v. 15.0 eggs/female) and in half-volume cages (3.9 v. 12.5 eggs/female/week). Under standard insectary conditions, mean weekly mortality-rates of P.papatasi females were 18.3 +/- 4.8% in October-November and 36.5-59.1% during the warmer months. None of the experimental conditions yielded any significant improvement in survival-rates.     

Inactivation of Single-Celled Ascaris suum Eggs by Low-Pressure UV Radiation

Intact and decorticated single-celled Ascaris suum eggs were exposed to UV radiation from low-pressure, germicidal lamps at fluences (doses) ranging from 0 to 8,000 J/m² for intact eggs and from 0 to 500 J/m² for decorticated eggs. With a UV fluence of 500 J/m², 0.44- ± 0.20-log inactivation (mean ± 95% confidence interval) (63.7%) of intact eggs was observed, while a fluence of 4,000 J/m² resulted in 2.23- ± 0.49-log inactivation (99.4%). (The maximum quantifiable inactivation was 2.5 log units.) Thus, according to the methods used here, Ascaris eggs are the most UV-resistant water-related pathogen identified to date. For the range of fluences recommended for disinfecting drinking water and wastewater (200 to 2,000 J/m²), from 0- to 1.5-log inactivation can be expected, although at typical fluences (less than 1,000 J/m²), the inactivation may be less than 1 log. When the eggs were decorticated (the outer egg shell layers were removed with sodium hypochlorite, leaving only the lipoprotein ascaroside layer) before exposure to UV, 1.80- ± 0.32-log reduction (98.4%) was achieved with a fluence of 500 J/m², suggesting that the outer eggshell layers protected A. suum eggs from inactivation by UV radiation. This protection may have been due to UV absorption by proteins in the outer layers of the 3- to 4-μm-thick eggshell. Stirring alone (without UV exposure) also inactivated some of the Ascaris eggs (~20% after 75 min), which complicated determination of the inactivation caused by UV radiation alone.     

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.     

UV irradiation of pig metaphase chromosomes: maturation-promoting factor degradation, nuclear cytology and cell cycle progression

Experiments were designed to test two hypotheses. The first was that irradiation of pig metaphase chromosomes would block the normal sequence of cytological and molecular events associated with activation; the second postulated that damaged DNA would prevent eggs from progressing through the first mitotic cleavage cycle. The experimental protocol involved selectively irradiating the metaphase II plate of pig oocytes with highly focused 254 nm ultraviolet (UV) light, followed by activation using standard electroactivation procedures. The following assessments were made of different groups of eggs: (i) nuclear membrane reassembly; (ii) chromosomal cytology; (iii) changes in maturation-promoting factor kinase (MPF kinase) activity at 1 h intervals after activation; and (iv) mitotic progression of eggs containing damaged chromosomal fragments. UV irradiation neither prevented the reassembly of nuclear membranes required for pronuclear formation nor interfered with the normal pattern of MPF kinase degradation after egg activation. UV irradiation did induce a wide range of chromatin defects, including condensation and dispersal of DNA fragments which, in turn, resulted in the formation of micronuclei in the treated eggs and embryos. The presence of damaged DNA retarded, but did not inhibit, progression through the first mitotic cycle. No evidence was obtained that the subsequent mitotic cycle was adversely affected by the presence of UV-damaged DNA. Overall, these results indicate that early cleavage divisions in pig eggs are not blocked by the presence of damaged, hypercondensed chromatin. In this respect, pig eggs are similar to Xenopus eggs, but are different from bovine eggs. On the basis of these findings it is suggested that focused UV irradiation offers a simple and rapid technique for the non-invasive enucleation of pig oocytes provided that the residual hypercondensed chromatin does not affect later developmental stages.     

Identification of mRNA-binding proteins during development: characterization of Bufo arenarum cellular nucleic acid binding protein

Ultraviolet irradiation was used to covalently cross-link poly(A)+RNA and associated proteins in eggs and embryos of the toad Bufo arenarum. Four major proteins with apparent sizes of 60, 57, 45 and 30-24 kDa were identified. It was observed that the same mRNA-binding proteins were isolated from eggs to gastrula and neural stages of development. The 30 kDa polypeptide, p30, appeared as the main ultraviolet (UV) cross-linked protein in the developmental stages analyzed. By means of polyclonal antibodies, it was determined that this polypeptide has a cytoplasmic localization and it was detected in liver, eggs and embryos. The presence of p30 was also analyzed by western blot during oogenesis and development. The 30 kDa polypeptide was present in all stages analyzed but it could not be detected in stages I-II of oogenesis. At the neural stage, the relative amount of p30 began to decrease, reaching its lowest levels after stages 26-30 (tail-bud in Bufo arenarum). On the basis of purification, immunoprecipitation and western blot assays the 30 kDa protein was identified as the Bufo arenarum cellular nucleic acid binding protein.     

Inhibition of cell growth by near ultraviolet light photoproducts of tryptophan

Exposure of dilute aqueous solutions of tryptophan to near UV light (320 to 390 nm) at subsolar levels yields fluorescent photoproducts capable of inhibiting the growth and differentiation of cultured mouse embryonic fibroblasts and fertilized sea urchin eggs. The ability of these cells to incorporate labelled precursors of protein, RNA, and DNA into their respective macromolecules was markedly inhibited by adding tryptophan preirradiated with near UV light to their incubation media. Thus the inhibition of growth and differentiation of these cells seems to result from a depression of their ability to synthesize macromolecules in the presence of the photoproducts.     

Restoration of fertility in sterilized Drosophila eggs by transplantation of polar cytoplasm

A technique for transplantation of cytoplasm between Drosophila eggs is described. Polar cytoplasm of newly laid eggs was first made ineffective for the determination of germ cells by UV irradiation. The sterility which results from this UV irradiation could be prevented by the injection of polar cytoplasm, but not by the injection of anterior cytoplasm from unirradiated donor eggs. The results provide the first demonstration of transplantation of agents causing determination in an insect and also provide a bioassay for these agents.Microscopical observations of UV irradiated eggs with and without transplanted polar cytoplasm revealed that UV irradiation delays the migration of cleavage nuclei into the posterior periplasm and prevents cytoplasmic protrusions at the posterior pole from becoming isolated from the periplasm to form pole cells. Transplantation of polar cytoplasm repairs these defects.     

Effects of selected physical and chemical treatments of Colorado potato beetle eggs on host acceptance and development of the parasitic wasp, Edovum puttleri

Effects of various physical and chemical treatments of Colorado potato beetle [Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)] eggs on parasitization and development of the egg parasitoid Edovum puttleri (Hymenoptera: Eulophidae) were investigated. UV irradiation did not affect host acceptance but reduced host suitability for UV exposure times 90 min. Susceptibility of host eggs to UV irradiation varied with host age; eggs were most vulnerable to damage from irradiation at 12, 18, and 24 h post-oviposition. The rate of parasitization also was influenced by host age. Percent parasitization was greatest in freshly laid eggs and 24–30 h old eggs. Seventy-seven percent of host eggs frozen at –20 °C (5 min) were parasitized by E. puttleri, but extended exposure of eggs to –20 °C reduced both acceptance and suitability. Host eggs that had been washed with hexane (removal of kairomone and sticky layer) also were parasitized. After 5 min of washing, application of kairomone significantly increased the rate of parasitism (from 74.7% to 88.2%), but with longer periods of washing, kairomone application had no significant effect on percent parasitism. Thus, the sticky material(s) coating the egg did not appear to be essential for parasitization to occur. Our results provide effective methods and times for treating Colorado potato beetle eggs to maximize parasitization and development of E. puttleri.     

泥鳅和大鳞副泥鳅卵雌核的紫外辐射灭活条件筛选

采用紫外线对泥鳅和大鳞副泥鳅卵雌性原核进行干法灭活以及在合成卵巢液、TC-199溶液、Ringer氏溶液和Holtfreter氏溶液中灭活,结果表明:当辐射剂量为90-180mJ/cm~2时,卵子在四种溶液中的单倍体诱导率均较高。合成卵巢液组的最佳辐射剂量为120-180mJ/cm~2;TC-199和Ringer氏溶液组的最佳辐射剂量为120mJ/cm~2;原液使用Holtfreter氏溶液效果欠佳,干法灭活效果最差。在上述各辐射剂量下,畸形苗经染色体组鉴定92％为雄核发育单倍体。综合评价,灭活效果依次是:人工合成卵巢液>TC-199溶液>Ringer氏溶液>Holtfreter氏溶液>干法。     

Storage of Euschistus heros Eggs (Fabricius) (Hemiptera: Pentatomidae) in Liquid Nitrogen for Parasitization by Telenomus podisi Ashmead (Hymenoptera: Platygastridae)

Records in the literature with regard to the influence of freezing of pentatomid eggs on parasitism by microhymenopterans are scarce. In this research, we compared the storage of Euschistus heros (Fabricius) (Hemiptera: Pentatomidae) eggs in liquid nitrogen for different periods with the objective of optimizing the multiplication of Telenomus podisi Ashmead (Hymenoptera: Platygastridae) in the laboratory. Fresh eggs of E. heros were exposed (S3, S6) or not (NS3, NS6) to UV light for 30 min and stored in 1.5-mL plastic vials in liquid nitrogen either for 3 (S3, NS3) or 6 months (S6, NS6), and egg suitability to parasitoid development was compared to control eggs exposed (SC) or not (NSC) to UV treatment. Global data analysis showed that E. heros eggs stored in liquid nitrogen with or without UV treatment, for 3 or 6 months, were suitable for T. podisi parasitization.     

Extreme tolerance to environmental stress of sexual and parthenogenetic resting eggs of Eucypris virens (Crustacea,Ostracoda)

1. The freshwater ostracod (Ostracoda), Eucypris virens, is commonly found in European temporary pools, where its long‐term persistence completely relies on the build‐up of resting egg banks. Extreme tolerance of dormant eggs and seeds is widely assumed, but freshwater ostracod eggs are relatively poorly studied. The study of ostracod resting eggs is of particular relevance as it may yield the key to understanding the distribution of the sexes in many species capable of both sexual and asexual reproduction. 2. We assessed the tolerance of dried resting eggs produced by females originating from three populations with males and three all‐female E. virens populations. Hatching time and success was compared between control eggs and eggs exposed to one of seven ecologically relevant stressors: digestive enzymes, high salinity, deep freezing, hydration, UV‐B radiation, hypoxia and insecticide treatment. 3. None of the stressors reduced significantly the viability of either sexual or asexual eggs. When compared with the reproductive mode–specific controls, exposure to UV‐B radiation had a mild impact on the survival of sexual and asexual eggs (?16.8 and ?22.4%, respectively), but this was only significant for asexual eggs. These results point to an extreme tolerance of E. virens resting eggs and have important implications for the ecology and evolution of the species. 4. The timing of hatching was not affected by the stress treatment, except for UV‐B radiation. A marginally significant delay in hatching response was observed for UV‐B‐radiated eggs when compared to the overall mean, but this treatment effect was absent when compared with the reproductive mode–specific controls. 5. The populations with males produced eggs that hatched on average earlier (?1.5 days at 17 °C) and were more successful (+26%) than asexual eggs. Due to the limited number of populations and the population‐specific origin and age of the eggs, the possibility due to the differences in age and origin of the resting eggs, or to variations in local conditions, cannot be ruled out.     

Immobile and tough versus mobile and weak: effects of ultraviolet B radiation on eggs and larvae of Manduca sexta

Although indirect effects of solar ultraviolet (UV) radiation on insects are well known (e.g. UV radiation can modify plant chemistry), direct effects of solar radiation on insects have received little attention. Radiation in the UVB range (300–320 nm) is damaging because it is absorbed directly by proteins and DNA. UVB should be toughest on immobile or small life stages, such as eggs or early larval instars. In the present study, the effects of UVB radiation on eggs and larvae of the tobacco hornworm Manduca sexta L. (Lepidoptera: Sphingidae) are examined. The present study aimed to address: what natural levels of UV do they experience; how does UVB affect the performance of eggs; and how does it affect the performance of larvae? In addition, do M. sexta larvae use behaviour to avoid UVB exposure and, consequently, are they physiologically less robust to UVB? In these experiments, eggs and late larval instars of M. sexta are found to be robust to natural levels of UV radiation. By contrast, young larvae are not only more susceptible to damage from UVB, but also they use behavioural means to avoid it. The strategy of using behaviour may relax selection pressures on morphological and physiological mechanisms for preventing (or recovering from) damage by environmental UV radiation.     

Ultraviolet and green parts of the colour spectrum affect egg rejection in the song thrush (Turdus philomelos)

Much attention has been devoted to understanding the evolution of egg mimicry in avian brood parasites. The majority of studies have been based on human perception when scoring the mimicry of the parasitic egg. Surprisingly, there has been no detailed study on the recognition and sensitivity towards differently coloured parasitic eggs. We investigated effect of different colours of the experimental eggs measured by ultraviolet (UV)-visible reflectance spectrophotometry on rejection behaviour in the song thrush ( Turdus philomelos ). We carried out a set of experiments with four blue model eggs representing mimetic eggs, whereas six other colours represented nonmimetic eggs. Our results revealed that two colours originally designed as a mimetic were rejected at a high rate, whereas one group of the nonmimetic was accepted. A multiple regression model of absolute differences between song thrush and experimental eggs on rejection rate showed that the level of mimicry in the UV and green parts of the colour spectrum significantly influenced egg rejection in the song thrush. To our knowledge, this is the first detailed study showing that different colour perception by the birds can affect their responses towards the parasitic egg. These findings suggest that the combination of UV and visible ranges of the spectra plays a major role in the evolution of discrimination processes, as well as in the evolution of the mimicry of the parasitic egg.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 269–276.