Intra-guild predation and variation in egg defence between sympatric and allopatric populations of two species of ladybird beetles

Abstract.  1. Ladybird eggs are defended chemically against intra-guild predation, and are unsuitable to varying degrees as food for other ladybirds.
2. Ladybird eggs [ Coccinella septempunctata (Cs) and Harmonia axyridis (Ha); Coleoptera: Coccinellidae] from local Japanese versus North American populations were compared as food for two ladybirds (Cs and Ha) co-occurring in Japan.
3. Larvae of Cs exhibited high mortality, slow development, and reduced egg consumption and weight gain when they fed on Ha versus conspecific eggs, especially when Ha eggs were from the local (Japanese) population versus a distant (North American) population of Ha.
4. Larvae of Ha survived equally well on a diet of Cs or conspecific eggs, but developed more slowly and gained less weight on Cs eggs. Ha larvae were more reluctant to eat eggs from the local (Japanese) population of Cs than eggs from the distant (North American) population of Cs; however, other measures of performance did not differ significantly.
5. These results indicate greater egg chemical defence of Ha eggs against Cs larvae than vice versa, as expected from field observations of greater temporal overlap between Cs larvae and Ha eggs than between Ha larvae and Cs eggs. Furthermore, results also indicate that local populations of eggs are better defended than eggs from elsewhere against locally occurring intra-guild predators. Thus, it appears that the effectiveness of chemical defence of ladybird eggs reflects the degree to which specific pairings of ladybirds have the potential to interact in nature through egg predation.     

The effect of embryonic development on metal and calcium content in eggs and eggshells in a small passerine

Published information relating to changes in the chemical element content of avian eggs caused by embryonic development is extremely scarce, although it may be crucial for understanding both the presence of anthropogenic pollutants as well as physiological levels of micronutrients. We assessed the variation in concentrations of calcium (Ca) and magnesium (Mg) and nine trace elements: seven essential (chromium (Cr), copper (Cu), nickel (Ni), manganese (Mn), iron (Fe), cobalt (Co) and zinc (Zn)) and two non‐essential (lead (Pb) and cadmium (Cd)) in shells and contents (both egg yolk and egg white) of embryonated and non‐embryonated eggs. We investigated the eggs of the Eurasian Reed Warbler Acrocephalus scirpaceus, a large proportion of whose eggs are infertile in our study population (almost 43% of clutches contain unhatched eggs) as well as significant embryo‐induced eggshell thinning at the equator of embryonated eggs. We found significantly higher concentrations (≥ 22.7%) of all the focal elements in the contents of embryonated eggs in comparison with non‐embryonated eggs, and a very pronounced one for Ca (nearly twice as high). The shells of embryonated eggs contained significantly higher concentrations of Zn (104.1%), Fe (56.5%), Pb (32.8%) and Cu (28.0%) but significantly lower ones of Co (8.9%) and Ca (9.3%) than the shells of non‐embryonated eggs. The simultaneous higher concentrations of all elements in the content of thinner‐shelled embryonated eggs suggest the parallel transfer of these elements along with Ca resorption from the shell into the egg interior during embryo formation. The higher concentration of most elements in the thinner shells of embryonated eggs may be indicative of the maternal deposition of some of these elements in a shell layer not subject to embryonic depletion, or in the eggshell membrane. Our results highlight the need for the careful selection of egg samples, which should differentiate between embryonated and non‐embryonated eggs in the analytical treatment of eggs and eggshells.     

Laboratory studies of Blattisocius keegani (Fox) (Acari: Ascidae) reared on eggs of navel orangeworm: potential for biological control

Blattisocius keegani (Fox) is a predatory mite in the family Ascidae (Acari), noted for potential biological control of Coleopteran stored product pests. Performance of B. keegani on eggs of navel orangeworm, Amyelois transitella Walker (Lepidoptera: Pyralidae), was investigated. Mites completed development from egg to adult in 9.2±0.22 days at 25°C and 50-60% relative humidity, and in 6.33±0.29 days at 32.2°C, 30% relative humidity. Mites provisioned with three or five eggs consumed a median of 1.25 to 1.5 eggs, with a maximum of three eggs consumed over 24?h. Regression analyses indicated egg-laying by B. keegani was significantly correlated with the number of A. transitella eggs consumed, and female mites laid an average of 5.82±0.44 eggs over 72?h. Blattisocius keegani, developed on fresh and frozen eggs, laid significantly more eggs when provided with fresh eggs (F3,26=6.16, P=0.0026) and were able to develop on frozen Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs as an alternative host. Mites were equally fecund when fed eggs stored at 0° or -20°C. Provisioning of adult moth bodies in addition to egg prey items increased mite fecundity, although it was demonstrated that B. keegani are phoretic on adult moths as well. The results are the first experimental evidence of B. keegani as a predator of Lepidopteran eggs, as a phoretic species, and of their potential for biological control of navel orangeworm.     

Differential gene expression during early embryonic development in diapause and non-diapause eggs of multivoltine silkworm Bombyx mori

Quantification of the differential expression of metabolic enzyme and heat-shock protein genes (Hsp) during early embryogenesis in diapause and non-diapause eggs of the silkworm B. mori was carried out by semi-quantitative RT-PCR. Data analysis revealed that, the phosphofructokinase (PFK) expression started at a higher level in the early stage (6 h after oviposition) in non-diapause eggs, while in diapause induced eggs, it started at a lower level. However, the PFK gene expression in diapause eggs was comparatively higher than in non-diapause eggs. PFK facilitates use of carbohydrate reserves. The lower level of PFK gene expression in the early stage of diapause induced eggs but comparatively higher level of expression than in non-diapause eggs is due to enzyme inactivation via protein phosphorylation during early embryogenesis followed by de-phosphorylation in later stage. The sorbitol dehydrogenase-2 (SDH-2) gene was down regulated in diapause induced eggs up to 24 h and its expression levels in diapause induced eggs coincided with that of PFK gene at 48h in non-diapause eggs. During carbohydrate metabolism, there is an initial temporary accumulation of sorbitol which acts as protectant. The down regulation of SDH-2 gene during the first 24 hours in diapause induced eggs was due to the requirement of sorbitol as protectant. However, since the diapause process culminates by 48 h, the SDH-2 gene expression increased and coincided with that of PFK gene expression. The trehalase (Tre) gene expression was at a lower level in diapause induced eggs compared to non-diapausing eggs. The induction of Tre activity is to regulate uptake and use of sugar by the tissues. The non-diapause eggs revealed maximum expression of GPase gene with major fluctuations as well as an overall higher expression compared to diapause induced eggs. The diapause process requires less energy source which reflects lower activity of the gene. Heat shock protein (Hsp) genes (Hsp20.4, 40, 70, and 90) revealed differential levels of expression in both the eggs at all stages of embryonic development. The present study thus provides an overview of the differential expression levels of metabolic enzyme and Hsp genes in non-diapause and diapause induced eggs of multivoltine silkworm B. mori within 48 h after oviposition, confirming the major role of in early embryogenesis.     

Bacterial Infection of Washed and Unwashed Eggs with Reference to Salmonellae

A survey of eggs in the Fraser Valley area of British Columbia was conducted to determine the level of bacterial contamination on washed and unwashed shell eggs and the incidence of Salmonella infection in shell eggs. Samples of eggs were taken from 15 grading stations. Determinations of eggs from 87 shipments showed that a higher proportion of eggs was heavily contaminated (>5-million organisms per egg) after washing than as received at the grading stations. This finding suggests the need for the development of more satisfactory washing procedures. No salmonellae were detected in the 180 samples (3,995 eggs) which were examined. It was concluded that Salmonella contamination of intact shell eggs does not constitute a serious health hazard in eggs from this area.     

Penetration of the zona-free or intact eggs by foreign spermatozoa and the fertilization of deer mouse eggs in vitro

Zona-free eggs were introduced to fresh or preincubated sperm suspensions and the penetration of eggs by foreign spermatozoa was examined, as evidenced by enlargement of the sperm head and formation of the male pronucleus. It was found that zona-free hamster eggs can be penetrated by guinea-pig, deer mouse and rabbit spermatozoa but zona-free rat, mouse and rabbit eggs cannot be penetrated by guinea-pig spermatozoa. Furthermore, zona-free rat and mouse eggs cannot be penetrated by spermatozoa from two species of deer mice and the Mongolian gerbil. The zona pellucida of a few intact rat eggs can be penetrated by mouse (6%) and by P. leucopus spermatozoa (14%) but enlargement of the sperm head and formation of pronuclei were observed in the former but not in the latter. It seems that (1) sperm capacitation is required for the penetration of zona-free eggs, (2) the attachment of foreign spermatozoa to eggs may indicate their potential ability of penetration in some cases, (3) there is a certain affinity between the vitellus of one species and spermatozoa from another species, (4) the block to the entry of foreign spermatozoa is not only in the zona pellucida but also in the vitelline membrane, (5) zona-free hamster eggs can be penetrated by spermatozoa of six species, (6) mouse spermatozoa can penetrate zona-free eggs of three species, and (7) fertilization of intact P. maniculatus eggs can be achieved in vitro.     

Egg-dumping lace bugs preferentially oviposit with kin

Egg dumping, or abandoning eggs and young to the care of other conspecifics, results in an extreme form of alloparental care. It is unclear, however, if egg dumpers discriminate among kin and nonkin egg recipients. In the lace bug Gargaphia solani (Heteroptera: Tingidae), some females with eggs (guards) also accept and defend eggs of conspecifics. Other females (egg dumpers) abandon their offspring after oviposition, leaving a single guard as the caregiver. We asked if egg dumpers preferentially dump their eggs among unguarded eggs of kin or nonkin. When given a choice between dumping among eggs of full siblings and eggs of nonsiblings, most eggs (67%) were dumped with full siblings' eggs. Furthermore, egg dumpers were just as likely to oviposit among eggs of kin with whom they had interacted on a shared host plant during juvenile development as they were to oviposit with kin reared on different host plants. Thus, egg dumpers discriminate kin by using cues associated with eggs, and such cues are not likely to be acquired through interaction on a common host plant environment. Copyright 2000 The Association for the Study of Animal Behaviour.     

Utilisation of lipids, protein, ions and energy during embryonic development of Australian oviparous skinks in the genus Lampropholis.

The contents of eggs and neonates of the Australian skinks, Lampropholis guichenoti and L. delicata, are described and compared to allow interpretation of nutrient utilisation by the developing embryo. Even though the females are the same size, L. guichenoti lay smaller clutches of larger eggs (egg contents=41.6+/-1.2 mg dry mass) than L. delicata (26.6+/-2.8 mg). The energy density is the same for eggs (30.5+/-0.9 J/g ash-free dry mass for L. guichenoti and 29.9+/-1.1 J/mg for L. delicata) and neonates (22.5+/-1.3 J/mg for L. guichenoti and 23.5+/-0.4 J/mg for L. delicata) between species. The amount of nitrogen (protein) in neonates is only slightly lower than that in eggs, whereas there is a large and significant decline in total lipids. Thus, like some other skinks, protein is a source of metabolic energy during embryogenesis, although not as important as lipid. Triacylglycerol is the major lipid component of the eggs (80% of total lipid), with phospholipid forming only approximately 10% of the total lipid. The fatty acid profile of the phospholipid is distinguished by a high proportion of arachidonic acid (8%), a significant proportion of eicosapentaenoic acid (2-4%) and a relatively low proportion of docosahexaenoic acid (2-3%) compared to chickens. Eggs of both species have remarkably low concentrations of free cholesterol compared to other amniote eggs (0.7% for L. guichenoti and 1.3% for L. delicata). The loss of lipid during embryonic development is almost entirely due to the selective utilisation of yolk triacylglycerol, presumably for energy. By contrast, the amount of phospholipid recovered from the neonates was the same as that originally in the eggs. Moreover, significantly more total cholesterol was present in the neonates than in the eggs, suggesting that biosynthesis of additional cholesterol occurred during development. The phospholipids of the neonates contain higher proportions of arachidonic (11-12%) and docosahexaenoic (8%) acids than the phospholipids of the eggs. Eicosapentaenoic acid is less prevalent in phospholipids in neonates than in eggs. Neonates of both species contain significantly more calcium than the fresh egg contents (L. guichenoti, eggs 0.303+/-0.051 mg, neonates 0.641+/-0.047 mg; L. delicata, eggs 0.187+/-0.013 mg, neonates 0.435+/-0.033 mg), presumably as a result of resorption of calcium from the eggshell. Interestingly, there is also significantly more sodium in neonates than in the contents of fresh eggs (L. guichenoti, eggs 0.094+/-0.010 mg, neonates 0.184+/-0.011 mg; L. delicata, eggs 0.084+/-0.011 mg, neonates 0.151+/-0.010 mg). There is no significant difference in the content of potassium and magnesium in eggs and neonates of either species. Although the fresh eggs of L. delicata have a significantly higher sodium concentration than L. guichenoti, there is no difference in the concentrations of calcium, magnesium, potassium or sodium in the neonates of the two species.     

Pronuclear location before the first cell division determines ploidy of polyspermic pig embryos.

Polyspermy occurs frequently in the fertilization of mammalian eggs, but little is known about whether polyspermic eggs have developmental ability in vitro or in vivo. We previously reported that poly-pronuclear (PPN; 3 or more pronuclei) pig eggs developed normally to the blastocyst stage despite having fewer inner cell mass cell numbers as compared to blastocysts derived from two-pronuclear (2PN) eggs. Here it is shown that most PPN pig eggs have abnormal cleavage patterns (having 3 or more cells) in the first cell division and retarded development of pronuclei prior to syngamy as compared to 2PN eggs. Most blastocysts (14 of 18) that developed from PPN eggs showed abnormal ploidy (were haploid, triploid, and tetraploid) whereas 20 of 22 blastocysts derived from 2PN embryos were diploid. The size and morphology of most Day 40 fetuses that developed from PPN eggs appeared to be normal. Of 8 Day 40 fetuses analyzed, 1 was triploid (XXY) and another was a mosaic with both diploid (XX) and tetraploid cells (frequency of less than 10%, XXXX), and the others were diploid. Anomalies of chromosomal composition were not detected in these fetuses. Five live piglets and one dead piglet were born from two recipients of PPN eggs. It is proposed that not all pronuclei of PPN pig eggs participate in syngamy, resulting in diploid cells in the conceptus. Our data suggest that there are two types of pronuclei location in polyspermic pig eggs and that the resulting ploidy is determined at the zygote stage before the first cell division according to pronuclear location.     

Effect of incubation and preservation on resting egg hatching and mixis in the derived clones of the rotifer Brachionus plicatilis

The marine rotifer Brachionus plicatilis typicus (Clone 8105A, Univ. of Tokyo) was cultured in 500 ml beakers to form resting eggs. Tetraselmis tetrathele was used as a culture food. Just after formation, resting eggs were exposed to various temperature (5–25 °C) and light regimes (24L: OD and OL : 24D). When eggs were exposed to light just after formation, the eggs hatched sporadically over a month. No hatching was observed for six months when eggs were preserved under dark conditions regardless of the temperature. These eggs hatched simultaneously after being exposed to light and eggs preserved at 5 °C showed twice as high hatching rate (40%) as that of eggs preserved at 15–25 °C (24%). Clones from resting eggs that were kept under different temperature and light regimes were reared individually to the third generation. Incubation at 25 °C with lighting produced the highest (5.4% and 5.2 %) rate of mictic females during their 2nd and 3rd generations, respectively. The lowest rates (0 and 1.5%) were found when the eggs were kept at 5 °C in total darkness for six months. A lower rate of amictic female production was found in clones with higher rates of mixis.     

Changes in FaRP-like peptide levels during development of eggs from the plant-parasitic cyst nematode, Heterodera glycines

The plant-parasitic cyst nematode Heterodera glycines requires a host plant to complete its life cycle, which involves hatching of infective juveniles that parasitize through root entry. A laboratory population of H. glycines grown on soybean, Glycine max, undergoes a sharp increase in maturity between 5 and 6 weeks in culture, as measured by the proportion of eggs containing well developed pre-hatch juveniles (late development eggs) versus eggs without visible juveniles (early development eggs). The median percent of eggs classified as late development, representing all samples taken from 4 to 7 weeks in culture, was 61%. For all samples taken up to 5 weeks, 80% scored below the median. In samples taken after 5 weeks, 15% scored below the median. This shift in population maturity was accompanied by a significant increase (P < 0.01) in the number of hatched juveniles present in each sample. There was also a significant increase (P < 0.02) in amount of FaRP-like peptide detected by specific ELISA. Total FaRP levels increased from 0.18 +/- 0.07 fMol FLRFamide equivalents per ng protein in early development eggs to 0.40 +/- 0.17 in late development eggs. The level remained high in hatched juveniles. HPLC/ELISA detected as many as nine potential FaRPs in H. glycines, two of which were specifically increased (P < 0.005) in hatched juveniles. The association of FaRPs with maturing eggs and the possible involvement of these neuropeptides with juvenile hatching and motility are discussed.     

枝角类休眠卵形成和萌发的研究进展

综述了多年来国内外对枝角类休眠卵的研究概况,重点介绍了休眠卵的形态、形成的条件、保存方式及萌发条件等.休眠卵通常呈圆形或椭圆形,外被一层厚的卵膜.休眠卵的形成主要依赖于生物因子(遗传因素、种群密度等)和非生物因子(温度、食物、光照等)的作用.影响休眠卵萌发的因子有休眠卵的保存方式、保存时间、萌发阶段的光照及温度等.     

Suitability of non-fertilized eggs of Homalodisca vitripennis for the egg parasitoid Gonatocerus morrilli

Gonatocerus morrilli (Howard) (Hymenoptera: Mymaridae) is an egg parasitoid used in California, USA to control glassy-winged sharpshooter (GWSS), Homalodisca vitripennis (Germar) (Hemiptera: Cicadellidae). Virgin GWSS females deposit non-fertilized eggs and mated females can exhaust sperm reserves for egg fertilization. However, nothing is known about Gonatocerus spp. performance when using non-fertilized GWSS eggs as hosts. Host age preference for oviposition and suitability of non-fertilized GWSS eggs as hosts for G. morrilli reproduction were investigated to determine whether non-fertilized eggs on sentinel plants could be used to monitor egg parasitoid populations. Gonatocerus morrilli parasitized all ages of GWSS eggs (1–8 days old) regardless if the host egg was fertilized or not. In choice tests (fertilized versus non-fertilized eggs), parasitoids failed to emerge as adults from non-fertilized eggs more often than from fertilized eggs. The results indicate that non-fertilized eggs were accepted by G. morrilli as suitable hosts for oviposition, but were less suitable for immature development compared to fertilized eggs.     

Modulation of spermatozoa and zona pellucida properties by the soluble acrosome reaction-inducing factor of the ovulated egg-cumulus complex

Three sources of hamster periovulatory fluids (± heat inactivation at 56°C), with bovine serum albumin (BSA) as control, were tested for effects on penetration of three classes of eggs by hamster sperm precapacitated in BSA. These fluids were a soluble extract of cumulus oophorus fluid (COF) from the ovulated hamster egg-cumulus complex, serum, and follicular fluid. Egg types were ovulated, salt-stored (ovulated), and follicular. In both COF and serum, there were significant differences among egg types in mean penetration, and significant effects of fluid addition. In contrast, there was no effect of follicular fluid and no differences between follicular and stored eggs. For the follicular eggs (combined data, normalized, ranked), patterns of response to the three factors (± heating) were different: only unheated COF and heated serum increased penetration significantly above BSA control levels (average rank 20.2, 41.4, 38, for BSA, COF (unheated), serum (heated), respectively). This indicated that the active component in COF was heat labile, not present in either serum or follicular fluid, and, therefore, of oviductal origin. Oviduct and/or COF exposure of eggs and sperm was tested for effects as an acrosome reaction inducing factor (ARIF) for acrosome reactions (AR; zonabound and free-swimming sperm) and on sperm:zona binding and penetration. The COF ARIF for free-swimming sperm AR was heat stable. Penetration of follicular eggs increased after incubation in COF prior to sperm addition, but a greater response occurred when COF was added to eggs with sperm. In kinetic experiments, 25 min following sperm attachment, follicular eggs had lost 41% of initially bound sperm, vs. 23% for ovulated eggs, and had only 16 AR sperm/egg, vs. 26 for ovulated. Follicular eggs incubated in COF (then washed three times) had the same number of bound AR sperm as ovulated eggs. Acid solubilized zona pellucida (ASZP) from ovulated eggs was more effective as an ARIF per zona than ASZP from follicular eggs. Zonae of follicular eggs, as eivdenced by dissolution times in β-mercaptoethanol (β-MEOH), were not “harder” than those of ovulated eggs. There were differences in lectin binding antigens on zonae of both fresh and stored, follicular and ovulated, eggs. We conclude that multiple biological factors orchestrate sperm:egg interactions in the ampulla. Our data are consistent with the presence of at least three effective components: (1) the oviductal lectin-binding antigen (ZPO or oviductin), (2) an additional heat-labile component, and (3) the heat-stable ARIF for free-swimming sperm. © 1994 Wiley-Liss, Inc.     

Identification of parasitized nests by using egg morphology in the Common Goldeneye: an alternative to blood sampling

We used data from three Common Goldeneye Bucephala clangula populations in Finland to study if the variation between females in egg morphology, as measured using a method developed by Eadie (1989), can be used to identify parasitized clutches. Eadie's method is based on z-score standardized measures of length, width, and weight of eggs. Using these measures, Euclidean distance for each pair of eggs within a clutch was calculated. Euclidean distance between the two most dissimilar eggs (maximum Euclidean distance, MED) was used as the criterion to identify parasitized clutches. Test clutches of 3 eggs that included one egg from each of three different females had a higher MED (2.80) than 3-egg clutches that included eggs from one female only (2.05), proving that there is statistically significant variation in egg morphology between females. Test clutches that included three eggs from each of three different females (9 eggs in all) had a mean MED of 4.51. The mean MED of naturally parasitized clutches (4.83) was higher than that of nonparasitized clutches (2.12). Further analyses suggested that MED>3.0 can be used as a conservative and reliable criterion to identify parasitized clutches. Our results confirm that Eadie's method is reliable enough to identify parasitized clutches in Common Goldeneyes.     

Consumption of flea faeces and eggs by larvae of the cat flea,Ctenocephalides felis

The effects of the consumption of flea faeces and non-viable eggs on larval development in the cat flea Ctenocephalides felis (Bouché) (Siphonaptera: Pulicidae) were investigated. Only 13.3% of larvae developed into adults when fed a diet of male or female flea faeces alone; however, 90% of larvae developed into adults when fed on flea faeces supplemented with non-viable flea eggs. When fed with non-viable eggs alone, larvae did not develop into adults. Nevertheless, non-viable eggs may provide critical supplemental nutrients, lacking in flea faeces and required for larval development. None of the larvae fed on flea faeces or non-viable eggs alone formed a cocoon. A diet of flea faeces alone significantly extended the second as well as third larval stadia compared to larvae fed on diets containing non-viable eggs. It is suggested that the cannibalism of fertile eggs may limit population growth in the cat flea.     

Cell Adhesion and Fusion Induced by Insemination in Zona-Free Hamster Eggs: Electrophysiological Monitoring of the Fusion Process

Cell adhesion and fusion were found to occur in zona-free hamster eggs placed in contact and then inseminated. A cytoplasmic fusion occurred in 7% of paired eggs mainly between 10 and 50 min after insemination. All other eggs that failed to fuse adhered tightly to one another within about 1 hr. Electrophysiological monitoring of the fusion process revealed that an electrical coupling (EC) suddenly appears between apposed eggs and completed within 2–15 min as the first step of cell fusion. Periodic hyperpolarizing responses (HRs), which have been found previously in fertilized hamster eggs to reflect a periodic increase in cytoplasmic Ca²⁺ ions, become gradually synchronized and completely coincide in paired eggs 10–30 min after the establishment of EC. Thus the paired eggs become an electrically single cell as the second step. Then the boundary between the eggs breaks down, resulting in formation of a single, spherical egg in several minutes. On the other hand, most of adhered eggs showed neither EC nor synchronization of HRs. Some adhered eggs showed only a low efficiency EC.     

Survival rates of parasite eggs in sludge during aerobic and anaerobic digestion.

The effects of mesothermic anaerobic or aerobic sludge digestion on survival of eggs from the roundworms Ascaris suum, toxocara canis, Trichuris vulpis, and Trichuris suis and from the rat tapeworm Hymenolepis diminuta were studied. Destruction of eggs throughout a 15-day treatment period, as well as their viabilities after reisolation, was analyzed. The laboratory model digesters used in this study were maintained at a 15-day retention schedule, partially simulating a continuously operating system. Ascaris eggs were destroyed in the anaerobic (23%) or aerobic (38%) digesters, and 11% Trichuris eggs were destroyed in the aerobic digesters. Trichuris eggs in anaerobic digesters and Toxocara eggs in either anaerobic or aerobic digesters were not destroyed. Destruction of eggs in digesters was correlated with the state of the eggs before subjection to the treatment processes; i.e., some Ascaris and Trichuris eggs were already embryonated in host intestinal contents or feces and hence past their most resistant stage. The viabilities of Ascaris and Toxocara eggs that survived the digestion processes were greater in anaerobically treated than in aerobically treated material. Eggs from Hymenolepis were nonviable before use in the experiments. However, they were more effectively destroyed in aerobic digesters than in anaerobic digesters.     

Establishment of self-sterility of eggs in the ovary of the solitary ascidian,Halocynthia roretzi

When unfertilized eggs (UFE) of the solitary ascidian, Halocynthia roretzi, are released naturally they are strictly self-sterile, whereas almost all ovarian eggs isolated after spawning are self-fertile. Self-sterile eggs are prepared within a relatively short period of several hours before the spawning. The morphological changes in ovarian eggs during late oogenesis were studied with special reference to the establishment of self-sterility. Four types of eggs at serial developmental stages were classified according to the morphology of their external envelopes. Self-sterility was established in the last stage, from the ovarian egg type 3 (OVE3) to UFE stages. Ovarian eggs which had become committed to UFE were denoted as full-grown ovarian eggs (FOE). FOE were able to differentiate into self-sterile UFE in vitro, whereas OVE3 could not. Several morphological differences between OVE3 and UFE were found. OVE3 had a germinal vesicle (GV), a type of vitelline coat (VC-OVE3) and no expanded perivitelline space, whereas UFE had completed germinal vesicle break down (GVBD), had another type of coat (VC-UFE) and showed an expanded perivitelline space. There were also some differences in the mode of fertilization between OVE3 and UFE. In UFE, sperm became bound firmly to the vitelline coat and passed through the coat with the help of follicle cells, whereas in OVE3, sperm did not bind so strongly and entered the perivitelline space without the aid of follicle cells. The relationships between the establishment of self-sterility and these morphological and functional changes in ovarian eggs are discussed.