Effects of pesticide applications on the euonymus scale (Homoptera: Diaspididae) and its parasitoid, Encarsia citrina (Hymenoptera: Aphelinidae)

Novel biorational insecticides are rapidly replacing more toxic, broad-spectrum compounds to control pests of ornamental plants. These new formulations are widely regarded as safe, effective, and environmentally sound with minimal impact on nontarget organisms. We tested several biorational and traditional insecticides for their ability to control euonymus scale, Unaspis euonymi (Comstock), and their potential impacts on the aphelinid parasitoid, Encarsia citrina (Crawford). Soil-applied acephate and foliar-applied pyriproxyfen exhibited superior control of euonymus scale, but also reduced numbers of surviving E. citrina. Imidacloprid failed to control euonymus scale and decreased parasitism by E. citrina. Thus, the potential impact of a pesticide on biological control is not necessarily predicted by its potential longevity, mode of delivery, or its toxicity to the target pest. Finding the best fit of a compound into an integrated pest management program requires a consideration of all these factors and direct study of effects on the natural enemies of pests.     

Development of Meteorus pulchricornis and regulation of its noctuid host, Pseudaletia separata

The solitary endoparasitoid Meteorus pulchricornis can parasitize many lepidopteran host species successfully. In the case of parasitization of Pseudaletia separata, developmental duration of M. pulchricornis was 8-9 days from egg to larval emergence and 6 days from prepupa to adult emergence. Successful parasitism by M. pulchricornis decreased with host age. Following parasitization of day-0 4th host instar, the parasitoid embryo, whilst still enclosed in serosal cell membrane, hatched out of the egg chorion 2 days after oviposition. Subsequently, the 1st instar parasitoid emerged from the surrounding serosal cell membrane. Serosal cells dissociated and developed as teratocytes 3.5 days after oviposition. One embryo of M. pulchricornis gave rise to approximately 1200 teratocytes, a number that remained constant until 6 days after parasitization, but decreased drastically to 200 at 7 days post-oviposition. The teratocytes of M. pulchricornis were round- or oval-shaped and grew from 65 microm at 4 days to 200 microm in the long axis at 6 days post-parasitization. At 4 days post-parasitization, many cells or cell clusters with lipid particles were observed in the hemocoels of parasitized hosts. In addition, paraffin sections of parasitized hosts revealed that many teratocytes were attached to the host's fat body and contributed to disrupting the fat body tissue. Further, examination of the total hemocyte count (THC) during parasitization revealed that THC was maintained at low levels. Surprisingly, a temporal decrease followed by restoration of THC was observed in hosts injected with virus-like particles of M. pulchricornis (MpVLPs) plus venom, which contrasts with the constant THC suppression seen in parasitized hosts. This indicates that MpVLP function is temporal and is involved in regulation of the host during early parasitism. Therefore, teratocytes, a host regulation factor in late parasitism, could be involved in keeping THC at a low level.     

Encarsia or Encarsiella? – redefining generic limits based on morphological and molecular evidence (Hymenoptera,Aphelinidae)

Abstract.  The genus Encarsiella ( syn.n. ) (Hymenoptera, Chalcidoidea, Aphelinidae) is synonymized with Encarsia on the basis of molecular and morphological characters. A phylogenetic analysis was conducted of the D2 expansion region of the 28S ribosomal DNA including thirty-eight Encarsia species. Our phylogenetic analysis revealed a polyphyletic origin of Encarsiella and confirmed the results of earlier analyses indicating the paraphyly of Encarsia . Four species are described as new ( Encarsia bifasciata sp.n. , E. caelata sp.n. , E. obliqua sp.n. and E. pilosa sp.n. ). The following species are transferred from Encarsiella to Encarsia : Encarsia aleurodici ( Girault, 1916 ) comb.n. , E. amabilis ( Huang & Polaszek, 1998 ) comb.n. , E. boswelli ( Girault, 1915 ) comb.n. , E. magniclava ( Girault, 1915 ) comb.n. , E. narroi Gómez & García comb.n. , E. nepalensis ( Polaszek & Hayat, 1992 ) comb.n. , E. noyesi ( Hayat, 1983 ) comb.n. , E. pithecura ( Polaszek, 1999 ) comb.n. , E. tachii ( Polaszek & Hayat, 1992 ) comb.n. , E. taiwanensis ( Chou & Chou, 1994 ) comb.n. and E. tamaulipeca ( Myartseva & Coronado-Blanco, 2002 ) comb.n. The generic limits of Encarsia are re-defined based on a re-assessment of morphological characters, and the results are discussed with respect to the current classification of the aphelinid subfamily Coccophaginae.     

桑白蚧的重要寄生蜂——桑白蚧恩蚜小蜂的鉴别与利用

桑白蚧恩蚜小蜂Encarsia(=Prospaltella)berlesel(Howard)是寄生桑白蚧Pseudaulacaspls pentagona(Targioni-Tozzctti)的重要寄生蜂,许多国家进行了引进移植,对控制桑白蚧的为害取得明显的成效．本记述了桑白蚧恩蚜小蜂形态特征的鉴别。以及各国引进利用的概况．并讨论了利用寄生蜂防治桑白蚧的重要性．     

Occurrence of G-banding in metaphase chromosomes of Encarsia berlesei (Hymenoptera: Aphelinidae).

Well-defined G-bands were obtained on somatic metaphase chromosomes of Encarsia berlesei using trypsin and warm 2x SCC in sequence. The G-banded pattern allowed rapid identification of all five metacentric chromosomes, which appeared uniformly lighted when stained with DAPI fluorochrome dye. It is stressed that ageing affects G-banding in this insect species; in fact, good banded chromosomes were obtained on 1-month air-stored chromosomes. Evidence for asynchronous condensation on the chromosomes of this species is also provided.     

Free serosal cells originating from the embryo of the wasp Diadromus pulchellus in the pupal body of parasitized leek-moth,Acrolepiosis assectella. Are these cells teratocyte-like?

In braconid species, teratocytes are derived from a serosal cell membrane which envelops the developing parasitoid embryo. On hatching, this membrane dissociates into individual cells, the teratocytes, which then circulate in the haemolymph of the host. We describe herein such a membrane, surrounding the embryo in eggs of the ichneumonid parasitoid wasp, Diadromus pulchellus. This membrane consisted of a single sheet of tightly packed cells with large 12+/-1.4 &mgr;m nuclei. These cells were released after hatching in vitro and cells of the same size were detected in vivo, in the vicinity of the D. pulchellus embryo. The number of nuclei detected suggests that the serosal membrane consists of about 450+/-150 cells. These cells did not grow after hatching of the parasitoid egg in the parasitized host, Acrolepiosis assectella, during the development of the parasitoid wasp larva. Southern blot experiments, using D. pulchellus satellite DNA or the ribosomal genes as probes, showed that free-living floating cells of wasp origin were present in the body of the parasitized host. This is the first time that free-floating teratocyte-like cells have been described in species of the Ichneumonidae.     

Developmental rates of two congeneric parasitoids,Encarsia formosa and E. pergandiella (Hymenoptera: Aphelinidae), utilizing different egg provisioning strategies

Endoparasitic Hymenoptera vary in the extent to which they provision their eggs and thus in the degree to which they appear to rely on their hosts for resources during embryonic development. In this study, developmental rates were examined in two congeneric parasitoid species, Encarsia formosa and E. pergandiella, that provision their eggs to different degrees. E. formosa eggs are much larger than E. pergandiella eggs. E. formosa eggs hatch significantly earlier than the eggs of E. pergandiella when deposited in 1st or 4th instar nymphs of a common whitefly host, Bemisia tabaci. Both species hatch earlier in 4th instar nymphs, but the delay in hatching in hosts parasitized as 1st instars is much greater in E. pergandiella. While E. formosa develops more rapidly to the 1st larval instar, E. pergandiella emerge as adults significantly earlier, though smaller, than E. formosa adults regardless of the host instar parasitized. These findings show that the extent of provisioning in the eggs of these wasps does not strictly determine their order of progression through different stages of development.     

Coexistence of two effective parasitoids of the white peach scale Pseudaulacaspis pentagona (Hemiptera: Diaspididae): the role of host stage and temperature

ENCARSIA BERLESEI: (Howard) and Pteroptrix orientalis (Silvestri) are endoparasitoids of Pseudaulacaspis pentagona (Targioni Tozzetti), that are sympatric in Campania (Italy). The influence of host stages on several components of E. berlesei fitness and the effect of mating status on the production of Pteroptrix orientalis progeny were evaluated at 25 degrees C. Parasitization of earlier host stages resulted in an increase in the development time and a decrease in progeny number of E. berlesei. Adult parasitoids were largest when young female stages were parasitized. Pupal mortality did not differ among host stages. Mated female P. orientalis produced a greater number of progeny and proportionally fewer sons (13.6% of the total progeny) than did virgin females (21.7%). The reproductive potential of both parasitoids was also evaluated at four regimes of constant temperature (20, 24, 26 and 30 degrees C). Encarsia berlesei attained rm values nearly double those of P. orientalis at 20, 24 and 26 degrees C, whilst at 30 degrees C the two parasitoids achieved the same rm value, since at this temperature E. berlesei suffered a high pupal mortality. Sex ratio of P. orientalis, expressed as proportion of males, varied significantly between 20 and 24 degrees C only.     

Growth patterns of Microplitis rufiventris (Hym., Braconidae) teratocytes in Spodoptera littoralis (Lep., Noctuidae) larvae treated with a chitin synthesis inhibitor

The influence of sublethal dietary levels (0.4 p.p.m.) of a chitin synthesis inhibitor lefenuron { N -[2,5-dichloro-4-(1,1,2,3,3,3-hexafluoropropoxy)-phenylaminocarbonyl]-2,6-difluorobenzamide} on the development of Microplitis rufiventris teratocytes was investigated. Observations and data were taken on maturing teratocytes, i.e. at the end of parasitoid development. When embryogenesis of the parasitoid eggs was initiated in treated hosts, the dissociated cells of the embryonic membrane of some of the parasitoid larvae did not liberate in the haemolymph of the host, whereas in the other hosts, the released ones as individual cells scored the largest mean number per host larva. When young teratocytes were exposed to the treatment they were found in some cases clumping in masses or clustered around the parasitoid larvae. In treated hosts, different size classes of teratocytes co-existed. Larger and morphological abnormal cells were more common in hosts which contained older teratocytes at the time of treatments. Abnormal teratocytes were associated with developmental abnormalities of parasitoid larvae, possibly due to deterioration of the host environment. This observation was common in treated hosts and in few cases of untreated ones. A significant decrease in teratocyte size occurred 24 h after parasitoid emergence. Observations and results obtained in the present work suggested that: (1) teratocyte may function in controlling the growth and successful development of parasitoid larvae; (2) they might have a protective function through sequestering abnormal materials issuing in the haemolymph of the host either during the course of parasitoid development or due to external stress; and (3) number of teratocyte cells liberated from the embryonic membrane of parasitoid egg is not constant.     

Cotesia kariyai teratocytes: growth and development

The growth and development of teratocytes was examined in the Cotesia kariyai-Pseudaletia separata system. Cotesia kariyai embryos released an average of 163 teratocytes at the time of hatching, 3.5 days after oviposition. The cells increased in diameter from 30 to 77 μm until 7 days post-parasitization, after which there was no significant increase in average diameter. However, there was significant variation in diameter within the population of teratocytes during the later developmental stages of the parasitoid larvae. The DNA contents increased up to day 7. The ploidy level of teratocytes increased 4-fold (2C to 8C) from days 4 to 7 and thereafter remained the same. Scanning electron microscopy (SEM) revealed that the surface of the teratocytes was covered with microvilli during all developmental stages, although on days 9 to 10 post-parasitization, bleb structures were also observed on a few. In vitro analysis of the proteins secreted from teratocytes following labeling with (35)S-methionine showed that many proteins were synthesized de novo and secreted by the cells until 9 to 10 days post-parasitization. These results indicate that teratocytes in later stages of development maintain their activity and regulate the physiological state of the host.     

Teratocytes of the solitary endoparasitoid Meteorus gyrator (Hymenoptera: Braconidae): morphology, numbers and possible functions

Abstract. Laboratory studies investigated the development of teratocytes derived from the eggs of the parasitoid Meteous gyrator (Thun.) in its host, the tomato moth Lacanobia oleracea (L.). At hatching, each parasitoid egg produced an average of approximately 1000 teratocytes, but this number declined to approximately 400 during the course of parasitism. The teratocytes increased in size markedly, such that 7 days after egg hatch their mean diameter was approximately four times that of the cells immediately after dissociation. The haemolymph of parasitized hosts had reduced phenoloxidase activity, and teratocytes inhibited phenoloxidase activity when coincubated with plasma from nonparasitized hosts. The injection of teratocytes into nonparasitized fifth‐instar L. oleracea larvae suppressed growth and induced a supernumerary moult in some larvae. A number of parasitism‐specific proteins were detected in the haemolymph of parasitized hosts, and incubation of teratocytes in culture media indicated that these cells were a source of at least two of these proteins.     

Morphometric and molecular analysis of the Encarsia inaron species-group (Hymenoptera: Aphelinidae), parasitoids of whiteflies (Hemiptera: Aleyrodidae)

Several series of host-reared specimens of an Encarsia species, initially thought to be the cosmopolitan Encarsia inaron (Walker), were collected in the Azores Islands (Portugal). Subsequent morphometric analysis supported the presence of two species: E. inaron and a new species, described herein as Encarsia estrellae Manzari & Polaszek sp. n. Encarsia estrellae was reared from Aleyrodes singularis Danzig, A. ?singularis, and Bemisia sp. afer-group on several host plants. In addition, the D2 region of the 28S rDNA gene was sequenced in eight individuals belonging to these species, as well as single representatives of two closely related and one distantly related species. Phylogenetic analysis of these DNA sequences, together with 23 additional Encarsia sequences retrieved from the European Molecular Biology Laboratory (EMBL) and GenBank databases, further supported the specific status of E. estrellae, and the placement of E. dichroa (Mercet) in the E. inaron species-group. Additionally, E. inaron is redescribed and some taxonomic problems in the E. inaron species-group are discussed.     

Extended in vitro culture of Microplitis croceipes teratocytes and secretion of TSP14 protein

Teratocytes, cells which originate from the serosal membrane of some Braconidae and Scelionidae, can be found in the hemocoel of permissive hosts during part or all of the developmental time of the parasitoid larva. Teratocytes from Microplitis croceipes are known to secrete biologically active proteins, which contribute to developmental arrest and failure to pupate of Heliothis virescens larvae. One such protein, which has a molecular weight of approximately 14 kDa is called TSP14. The presence of parasitoid larvae is essential to maintain teratocytes under in vitro conditions with protein-free EX-CELL 400. The teratocyte viability was maintained in vitro for at least 12 days in the presence of larvae when medium was exchanged every three days. Western blots show that TSP14 was secreted during the entire period of exchanges. In the absence of parasitoid larvae, teratocyte viability was only 30% by day 6 and no TSP14 had been secreted. In the absence of parasitoid larvae, teratocytes maintained in vitro in EX-CELL 400 medium supplemented with 10% FBS remained viable for at least nine days and secreted TSP14 for at least six days. This suggests that parasitoid larval secretions are sufficient but not uniquely essential to maintain teratocyte viability. Parasitoid larvae maintained in the absence of teratocytes did not secrete TSP14 and their secretory products did not inhibit pupation of H. virescens larvae.     

Additive effect of teratocyte and calyx fluid from Cotesia plutellae on immunosuppression of Plutella xylostella

Abstract.  Teratocytes are cells that originate from the extra-embryonic tissues of some hymenopteran parasitoids, typically dissociate upon hatching, and develop in the host haemolymph. They are considered to be involved in parasitoid larval nutrient uptake, host immunosuppression and/or repression of competing parasitoid development. Teratocytes of the parasitoid, Cotesia plutellae (Kurdjumov) (Hymenoptera: Braconidae) are found in its natural host, Plutella xylostella (Linnaeus) (Lepidoptera: Yponomeutidae) and can be cultured in vitro . The present study demonstrates that teratocytes of C. plutellae possess a significantly depressive effect on host cellular immunity. When the hosts are preinjected with 200 cultured teratocytes (corresponding to the normal number of teratocytes released during wasp hatching), haemocyte nodulation is inhibited by approximately 40%, with younger teratocytes being more potent than older ones. Similarly, the medium in which teratocytes are cultured has similar immunosuppressive properties. In comparison, calyx fluid extracted from the C. plutellae ovary also has an immunosuppressive effect on P. xylostella . These two maternal (calyx fluid) and embryonic (teratocytes) factors are additive and result in a reduced level of nodule formation equivalent to that induced by natural parasitization. However, the immunosuppression of the parasitized P. xylostella does not appear to be due to inhibition of phospholipase A₂, an immune mediator, because injection of arachidonic acid failed to restore haemocyte nodulation capability.     

Interspecific host discrimination and within-host competition between Encarsia formosa and E. pergandiella (Hymenoptera: Aphelinidae), two endoparasitoids of whiteflies (Hemiptera: Aleyrodidae)

Interspecific host discrimination and within-host competition between Encarsia formosa Gahan and Encarsia pergandiella (Howard), two endoparasitoids of whiteflies, were studied under laboratory conditions. Interspecific host discrimination was studied at two time intervals (0 h and 72 h after the first species had oviposited). Parasitized and unparasitized Trialeurodes vaporariorum (Westwood) hosts were accepted for oviposition at the same rate by the two parasitoid species. Host type did not affect the handling time of the two parasitoids. The outcome of within-host competition was investigated after females of the two species parasitized the hosts at various time intervals. In four treatments, E. pergandiella was allowed to oviposit 0, 24, 48 and 72 h after E. formosa while in the other two, E. formosa was allowed to oviposit 0 and 72 h after E. pergandiella. In four of these treatments: E. formosa following E. pergandiella at 0 and 72 h, and E. pergandiella following E. formosa at 0 and 24 h, E. pergandiella prevailed. In the host discrimination experiment (72 h interval), 20% of E. pergandiella eggs were killed by E. formosa females. Interspecific ovicide was also observed in the within-host competition experiment, in which 6% of 72-h-old E. pergandiella eggs were killed by E. formosa females.     

Heterospecific ovicide influences the outcome of competition between two endoparasitoids, Encarsia formosa and Encarsia luteola

Abstract.  1. Studies of inter-specific competition in parasitoids have largely focused on the outcome of within-host competition and the behavioural mechanisms by which female parasitoids prevent competition. Another, less well studied, possibility is oviposition preceded by 'heterospecific ovicide', the destruction of the other species' egg. Heterospecific ovicide essentially eliminates within-host competition.
2. This study investigated the mechanisms and outcome of within-host competition in Encarsia formosa and Encarsia luteola , solitary endoparasitoids of whitefly pests. These species are known to commit ovicide of conspecific eggs.
3. Competition experiments indicated that the offspring of second-ovipositing females had an apparent advantage in competition, regardless of whether the second female was E. formosa or E. luteola .
4. Observations of ovipositor movement through the cuticle of host whitefly nymphs showed that both species often committed heterospecific ovicide and then oviposited or host-fed. Multiparasitism and heterospecific host discrimination were less common and absent respectively.
5. Heterospecific ovicide appears to explain the second-female advantage in competition between these species. Second-female advantage is contrary to the paradigmatic view of multiparasitism, where the first-ovipositing female has an advantage in competition or one of the species consistently prevails in competition.     

Host-parasite interactions between whiteflies and their parasitoids

There is relatively little information available concerning the physiological and biochemical interactions between whiteflies and their parasitoids. In this report, we describe interactions between aphelinid parasitoids and their aleyrodid hosts that we have observed in four host-parasite systems: Bemisia tabaci/Encarsia formosa, Trialeurodes vaporariorum/E. formosa, B. tabaci/Eretmocerus mundus, and T. lauri/Encarsia scapeata. In the absence of reported polydnavirus and teratocytes, these parasitoids probably inject and/or produce compounds that interfere with the host immune response and also manipulate host development to suit their own needs. In addition, parasitoids must coordinate their own development with that of their host. Although eggs are deposited under all four instars of B. tabaci, Eretmocerus larvae only penetrate 4th instar B. tabaci nymphs. A pre-penetrating E. mundus first instar was capable of inducing permanent developmental arrest in its host, and upon penetration stimulated its host to produce a capsule (epidermal in origin) in which the parasitoid larva developed. T. vaporariorum and B. tabaci parasitized by E. formosa initiated adult development, and, on occasion, produced abnormal adult wings and eyes. In these systems, the site of parasitoid oviposition depended on the host species, occurring within or pressing into the ventral ganglion in T. vaporariorum and at various locations in B. tabaci. E. formosa's final larval molt is cued by the initiation of adult development in its host. In the T. lauri-E. scapeata system, both the host whitefly and the female parasitoid diapause during most of the year, i.e., from June until the middle of February (T. lauri) or from May until the end of December (E. scapeata). It appears that the growth and development of the insects are directed by the appearance of new, young foliage on Arbutus andrachne, the host tree. When adult female parasitoids emerged in the spring, they laid unfertilized male-producing eggs in whiteflies containing a female parasitoid [autoparasitism (development of male larvae utilizing female parasitoid immatures for nutrition)]. Upon hatching, these male larvae did not diapause, but initiated development, and the adult males that emerged several weeks later mated with available females to produce the next generation of parasitoid females. Thus, the interactions that exist between whiteflies and their parasitoids are complex and can be quite diverse in the various host-parasitoid systems.     

A revision of the Encarsia longifasciata species group (Hymenoptera: Aphelinidae)

Abstract.  The Encarsia longifasciata group is established and revised, and its relationship to other species groups of Encarsia is discussed with reference to biological, morphological and molecular evidence. Principal component analysis on all available specimens provided supporting evidence for the existence of five species, two of which are described herein as new: E. dewa Pedata & Polaszek, sp.n and E. prinslooi Pedata & Polaszek, sp.n. All species are either diagnosed or described, and illustrated, and information is given on their distribution and host range. Additional information on the biology and larval stages, and scanning electron micrographs of morphological details, are provided for E. arabica Hayat. A preliminary phylogenetic study using 28S D2 rDNA sequence data for E. arabica was inconclusive, but suggested possible affinities between the longifasciata and parvella species groups. An illustrated dichotomous key to all species of the E. longifasciata group is given.     

Competition, resource partitioning and coexistence of an endoparasitoid Encarsia perniciosi and an ectoparasitoid Aphytis melinus of the California red scale

Abstract. 1. Laboratory experiments and field studies were conducted to explain the coexistence of an endoparasitoid, Encarsia perniciosi Tower, and an ectoparasitoid, Aphytis melinus DeBach, both of which were introduced into California to control the California red scale, Aonidiella aurantii (Mask.).
2. Encarsia parasitized all scale stages but it preferred first and second instar scales. This is in contrast to Aphytis melinus , in which previous studies have shown that it parasitizes second and third instar females and second instar males but prefers third instar female scales. Encarsia developed most rapidly when it parasitized an early second instar and slowest when it parasitized the mature female scale. However, on early second instar scales it was about 80% as fecund as a wasp that emerged from a mature female scale.
3. Second instar scales parasitized by Encarsia were accepted by Aphytis as readily as unparasitized scales.
4. Encarsia did not distinguish between unparasitized hosts and those previously parasitized by Aphytis.
5. Encarsia always outcompeted by Aphytis when both species parasitize the same host.
6. Encarsia prefers scale on stems whereas Aphytis prefers those on leaves and fruits. This, too, may be a result of interspecific competition with Aphytis.
7. The partitioning of the scale resource by the two species explains why they coexist in coastal southern California but it does not explain why Encarsia disappeared from citrus groves in the inland valleys coincident with the introduction of Aphytis melinus into southern California.     

Ultrastructure of Microplitis croceipes (Cresson) (Braconidae : Hymenoptera) teratocytes

The developing embryo of the braconid, Microplitis croceipes (Braconidae : Hymenoptera), is encased in an extraembryonic serosal membrane. Hatching of the parasitoid within the larva of its habitual host, Hehothis virescens (Noctuidae : Lepidoptera), is initiated about 40 hr after oviposition when held at 25 ± 2°C. At this time, the monolayered serosal membrane begins to dissociate into individual cells (teratocytes). After dissociation, teratocytes become dispersed in the hemolymph of the host. The average number of teratocytes released from each parasitoid embryo is 914 ± 43. Teratocytes average 14.1 ± 2.4 μm in diameter when first released, and reach a maximum average diameter of 68.1 ± 4.6 μm 6 days after liberation. Newly released teratocytes have ovoid nuclei, simple mitochondria and a limited number of profiles of the endoplasmic reticulum, all of which indicate relative metabolic inactivity. The ramified nuclei, extensive endoplasmic reticulum, polymorphic mitochondria and accumulation of glycogen granules and lipid droplets observed in older teratocytes provide circumstantial evidence that protein synthesis is occurring. Within hours after dissociation, microvilli begin to cover the surface of the teratocytes. Anatomical deformation (blebs) that occurred on some older (8-day-old) teratocytes probably resulted from enlargement or expansion of microvilli.