Evasion of encapsulation by parasitoid correlated with the extent of host hemocyte pathology

The egg and larval stages of the generalist endoparasitoid Campoletis sonorensis Carlson (Hymenoptera: Ichneumonidae), which normally avoid the hemocytic reaction of many Lepidopteran host species, are encapsulated in 40% of Spodoptera frugiperda J. E. Smith (Lepidoptera: Noctuidae) larvae. The effect of parasitism on inhibiting the spreading ability of S. frugiperda plasmatocytes in vitro is more pronounced in susceptible larvae which fail to encapsulate the parasitoid than in resistant ones permitting parasitoid development. This suggests that induction of plasmatocyte pathology is relevant to the successful evasion of encapsulation by the parasitoid. Some granular cells disappear from the hemolymph of the parasitized resistant larvae, which implicates their involvement in the encapsulation reaction. Calyx fluid of C. sonorensis injected into host larvae produced effects on host hemocytes identical to natural parasitism. Several mechanism may cooperate to protect the parasitoid from encapsulation. The pathological reactions by the host plasmatocytes is one main manifestation of the immunosuppressive parasitoid effect. Results are discussed in regard to the known effects of C. sonorensis on Heliothis virescens Fabricius (Lepidoptera: Noctuidae) larval hemocytes which are totally unable to respond with a successful cellular defense reaction.

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Résumé L'ichneumonide Campoletis sonorensis Carlson, endoparasitoïde larvaire, se développe dans de nombreuses espèces de Lépidoptères. Son statut de généraliste est dû, notamment, à son aptitude à déjouer les défenses immunitaires de ses hôtes, c.a.d. la formation d'une capsule d'hémocytes autour de l'oeuf ou de la larve parasite. Cependant, chez le noctuide Spodoptera frugiperda J.E. Smith, C. sonorensis est encapsulé dans 40% des larves qu'il parasite. Nous avons étudié la population hémocytaire de deux catégories de larves de S. frugiperda, celles qualifiées de résistantes à C. sonorensis, et qui arrêtent son développement, et celles susceptibles où le parasitoïde échappe à l'encapsulation.Cinq types d'hémocytes ont été identifiés: les prohémocytes (PR), les sphérulocytes (SP), les granulocytes (GR), les plasmatocytes (PL) et les oenocytoïdes (OE). Chez les hôtes susceptibles et résistants, le parasite provoque une baisse identique de la concentration totale des hémocytes dans l'hémolymphe (THC). Par contre, les PLs sont davantage affectés chez les hôtes susceptibles que chez ceux résistants au parasitoïde. Les résultats montrent que, chez les hôtes susceptibles, 1) le nombre des PLs dans l'hémolymphe est davantage diminué, et 2) leur aptitude d'adhérence in vitro est davantage inhibée. Il existe donc une corrélation positive entre le degré de pathologies qui affectent les PLs de l'hôte et l'incapacité de celui-ci à encapsuler le parasitoïde. Ceci tend à démontrer le rôle-clé des PLs dans la réaction immunitaire d'encapsulation chez S. frugiperda, comme chez de nombreux insectes. De plus, ce résultat renforce l'hypothèse selon laquelle C. sonorensis éviterait l'encapsulation en agissant sur les hémocytes de l'hôte, et plus particulièrement sur les PLs. Inversement aux PLs, les GRs sont moins abondants dans l'hémolymphe des hôtes qui encapsulent C. sonorensis. Les GRs pourraient donc participer à la formation de la capsule hémocytaire.Il est possible que plusieurs facteurs contribuent à protéger C. sonorensis de l'encapsulation. Néanmoins, les pathologies affectant les hémocytes des hôtes parasités sont probablement une manifestation majeure de l'effet immunosuppresseur du parasitoïde.Les effets de C. sonorensis sur les hémocytes des larves parasitées peuvent être reproduits chez des larves saines, en leur injectant de venin extrait des glandes du calyx des femelles parasitoïdes. Ces sécrétions provenant de la glande du calyx, et normalement injectées dans l'hôte lors de l'oviposition, sont probablment responsable, au moins en partie, de l'effet immunosuppresseur du parasitoïde.Ces résultats peuvent être comparés à ceux obtenus chez l'hôte Heliothis virescens Fabricius (lépidoptère, noctuide) qui n'encapsule jamais C. sonorensis. Bien que le parasitoïde provoque les mêmes effets pathologiques sur les hémocytes des hôtes de S. frugiperda et d'H. virescens, on constate que l'effet apparait quelques heures après l'oviposition chez H. virescens, contre 48 heures post-oviposition chez S. frugiperda. Ce délai pourrait contribuer à la résistance immunitaire de certaines larves de S. frugiperda, résistantes, à C. sonorensis.

     

Unaltered hematocrit values and differential hemocyte counts in acanthocephalan-infected Armadillidium vulgare

Three cell types were present in Armadillidium vulgare hemolymph. Hyaline cells, 8.2 to 12.0 μm in size with a few fine granules in the cytoplasm, comprised 42% of the cells. Twelve percent of the cells were nonexplosive granulocytes, 8.8 to 15.0 μm, with many cytoplasmic granules of medium size. A third cell type, an explosive 7.2- to 12.0-μm granulocyte with coarse cytoplasmic granules, comprised 46% of the cells. Within 48 hr after adult specimens of A. vulgare were fed eggs of the acanthocephalan, Plagiorhynchus cylindraceus, isopod hemocytes aggregated in the intestinal epithelium surrounding penetrating acanthors. Encapsulation and death of the parasite routinely followed. No significant difference in hematocrit values or in differential hemocyte counts occurred between infected and uninfected control isopods.     

N-acetyl-D-glucosamine in crustacean hemocytes; possible functions and usefulness in hemocyte classification

Abstract. The lectin wheat-germ agglutinin (WGA) selectively binds N-acetyl-D-glucosamine. Fluorescence and electron microscopy were used to show that WGA stains the cytoplasmic granules in the granulocytes, but not the hyaline cells, of two decapods, the ridgeback prawn Sicyonia ingentis and the American lobster Homarus americanus. Using fluorescence microscopy, two intermediate stages in granulocyte maturation were observed. Cells smaller than typical small-granule hemocytes were observed with 5 or fewer granules, which in previous studies using brightfield and phase optics were probably counted as hyaline cells. Also, some granulocytes were observed containing both small and large granules, supporting the suggestion that small and large granule hemocytes represent stages in the maturation of one cell line. Granules in the single type of hemocyte in the branchiopod Artemia franciscana did not stain with WGA. The possible roles of N-acetyl-D-glucosamine in wound healing, pathogen encapsulation, and maintenance of normal crustacean connective tissues are discussed.     

Freeze-fracture studies on the cockroach hemocyte membrane complex: Symmetric and asymmetric membrane particle distributions

Summary Freeze-fracture studies were conducted on the membranes of normal cockroach hemocytes. The plasmalemma is asymmetric with the A fracture face containing 80–100 Å membrane intercalated particles at a concentration of 2500/². The B fracture face contains 120–150 Å particles with a relatively low density (800/²). The nuclear envelope displays an asymmetry with the A fracture face containing 1500 particles/² and the B face containing 300/ ². No significant particle size differences were observed in nuclear envelope fracture faces. Two types of symmetric membranes were also found in these cells. Both A and B fracture faces of the membrane surrounding the numerous cytoplasmic inclusion bodies contain particle sizes and concentrations similar to the B face of the plasmalemma. A second type of symmetry was observed in cells apparently engaged in exocytosis. Vesicles (0.1 D) from this process were completely particle free on both fracture faces. Such particle free vesicles could be found in the cytoplasm, attached to the plasmalemma, or completely separated from the cell.Supported by a Pharmaceutical Manufacturers Association Foundation Fellowship.The author wishes to thank Ms. Annalena K. Charla for assistance in plate preparation, Dr. Julius Schultz and the Papanicolaou Cancer Research Institute for use of the freeze-etch device, and Dr. David Smith for the electron microscope facilities.     

敲低piwi基因对黑腹果蝇血细胞增殖及分化的影响

[目的]探究敲低piwi基因对黑腹果蝇Drosophila melanogaster血细胞增殖及分化的影响.[方法]利用黑腹果蝇e33C-Gal4和Hml-Gal4-UAS-2×EGFP品系分别与野生型W1118和UAS-piwi RNAi品系杂交,实现在黑腹果蝇游离血细胞或淋巴腺中降低piwi基因的表达;采用免疫荧光...     

Polygenic traits and parasite local adaptation

The extent to which parasites are locally adapted to their hosts has important implications for human health and agriculture. A recently developed conceptual framework--the geographic mosaic theory of coevolution--predicts that local maladaptation should be common and largely determined by the interplay between gene flow and spatially variable reciprocal selection. Previous investigation of this theory has predominately focused on genetic systems of infection and resistance characterized by few genes of major effect and particular forms of epistasis. Here we extend existing theory by analyzing mathematical models of host-parasite interactions in which host resistance to parasites is mediated by quantitative traits with an additive polygenic basis. In contrast to previous theoretical studies predicated upon major gene mechanisms, we find that parasite local maladaptation is quite uncommon and restricted to one specific functional form of host resistance. Furthermore, our results show that local maladaptation should be rare or absent in studies that measure local adaptation using reciprocal transplant designs conducted in natural environments. Our results thus narrow the scope over which the predictions of the geographic mosaic theory are likely to hold and provide novel and readily testable predictions about when and where local maladaptation is expected.     

Hemolymph melanization and alterations in hemocyte numbers in Lymantria dispar larvae following infections with different entomopathogenic microsporidia

Lymantria dispar (L.) (Lepidoptera: Lymantriidae) larvae can be infected in the laboratory with a variety of entomopathogenic microsporidia. In many cases, however, L. dispar is only a semi‐permissive host for such infections. In this study, we analyzed changes in the melanization of hemolymph and hemocyte numbers in L. dispar larvae after inoculation with various entomopathogenic microsporidia. We compared the infections produced by microsporidia isolated from L. dispar and infections produced by isolates from other Lepidoptera to which L. dispar is only a semi‐permissive host. Microsporidiosis induced a significant activation of the prophenoloxidase system leading to melanization; activation was highest when the pathogen caused heavy infections of the fat body, which was the case with two microsporidia originally isolated from L. dispar. Infection of only the silk glands or light infection of the fat body by two Vairimorpha spp. from other lepidopteran hosts elicited a lower response. Very light infections caused by a microsporidium isolated from Malacosoma americanum were not accompanied by elevated hemolymph melanization activity. Heavy infections by Endoreticulatus spec. that remained restricted to the gut tissue likewise did not elicit melanization. One Vairimorpha spec. from L. dispar induced a significant increase in total hemocyte numbers; the other infections led to temporarily decreased numbers. Microscopic examinations showed that parts of infected tissue were encapsulated by hemocytes. We conclude that measured alterations in hemolymph melanization and hemocyte numbers were likely to be induced by the damaging effects of heavy infections. Observed defense responses did not prevent the progression of infections.     

昆虫细胞免疫反应中的吞噬、集结和包囊作用

细胞免疫是昆虫天生免疫系统中很重要的部分, 包括了由血细胞介导的一系列吞噬、 集结和包囊等作用。本文讨论了近年来在昆虫细胞免疫方面的研究进展, 包括参与昆虫细胞免疫的血细胞类型, 识别外来异物的受体因子, 影响免疫活性的一些酶和化学物质等。另外还就吞噬模式, 以及集结和包囊过程中粘附态细胞的形成等加以讨论。     

Elevation of aminopeptidase activity in Biomphalaria glabrata (Mollusca) parasitized by Echinostoma lindoense (Trematoda)

Comparisons of the levels of aminopeptidase activity in the hemocytes and serum of Biomphalaria glabrata at 20 and 30 days postexposure to irradiated Echinostoma lindoense miracidia with enzyme levels in control snails have revealed that there are significant elevations in the serum of snails at both time periods postexposure. Furthermore, there is a significantly higher level of aminopeptidase activity in the serum of snails at 30 days than at 20 days postexposure. Although the biologic function of the elevated levels of serum aminopeptidase in sensitized snails remains uncertain, it is possible that this lysosomal enzyme may degrade the surface proteins of secondarily introduced parasites and thus act as a form of acquired humoral immunity.     

Certain hemocyte proteins of the medfly,Ceratitis capitata,are responsible for nonself recognition and immobilization of Escherichia coli in vitro

The results indicate that certain hemocyte proteins of the medfly, Ceratitis capitata, are responsible for the recognition of foreignness, since they are able to bind to the surface of Escherichia coli in vitro. Furthermore, when the E. Coli-hemocyte protein complex was incubated in the presence of tyrosine and phenoloxidase, the bacteria were immobilized, forming large aggregates. The formation of aggregates seems to be due to reactive tyrosine intermediate (quinone) generated by the action of phenoloxidase. © 1992 Wiley-Liss, Inc.     

Purification and binding properties of hemagglutinin from Biomphalaria glabrata.

A hemagglutinin has been purified from Biomphalaria glabrata (PR-B) hemolymph, albumin glands, and egg masses using affinity chromatography with Sephadex gels. The purified material from any of the sources above demonstrated identical immunological properties during immunoelectrophoresis or immunodiffusion, and similar serological specificity for human A₁ erythrocytes and to a lesser extent A₂ erythrocytes. Hemagglutinin was able to bind in vitro to the tegumental surface of cultured Schistosoma mansoni sporocysts, cercariae, and miracidia. Sporocysts dissected from infected snails and shed cercariae were already found to have hemagglutinin on their tegumental surface as demonstrated by immunofluorescence. It is postulated that hemagglutinin binding to the surface of larval helminths may “mask” them from being recognized by the snail host's cellular defense system.     

A secretory PLA2 associated with tobacco hornworm hemocyte membrane preparations acts in cellular immune reactions

We report on a secretory phospholipase A2 (sPLA2) associated with membrane-enriched fractions prepared from hemocytes of the tobacco hornworms, Manduca sexta. Virtually no PLA2 activity was detected in serum of immunologically naive or bacterially challenged hornworms. PLA2 activity was detected in cytosolic and membrane-enriched fractions prepared from hemocytes. PLA2 activity in the cytosolic fraction (1.2 pmol/mg/h) was approximately 4-fold greater than in the membrane-enriched fraction. The cytosol-associated PLA2 activity was strongly inhibited in reactions conducted in the presence of the specific cytosolic PLA2 inhibitor methylarachidonyl fluorophosphate (MAFP) but not in the presence of the sPLA2 inhibitor p-bromophenacyl bromide (BPB). Conversely, the membrane-associated PLA2 activity was inhibited in reactions conducted in the presence of BPB but not in the presence of MAFP. While the cytosol-associated PLA2 was independent of calcium, the membrane-associated sPLA2 required calcium for full catalytic activity. Hornworms treated with either BPB, MAFP or the glucocorticosteroid dexamethasone were severely impaired (by 50 to 80% relative to controls) in their ability to form nodules in reaction to bacterial challenge. However, the immune-impairing influence of the inhibitors was reversed by treating larvae with arachidonic acid, a precursor for eicosanoid biosynthesis. We infer that the biological significance of the sPLA2 (as well as the previously characterized cytosolic PLA2) relates to hydrolysis of polyunsaturated fatty acids from cellular phospholipids. Moreover, this enzyme may be the target of immunity-impairing factors from the bacterium Xenorhabdus nematophila. The fatty acids serve as precursors for the generation of eicosanoids responsible for mediating and coordinating cellular immune reactions to infection.     

Evidence for a LPS-binding protein in medfly hemocyte surface: mediation in LPS internalization but not in LPS signaling

A doublet of medfly hemocyte proteins with a molecular mass of about 55 and 50 kDa were precipitated with LPS. Antibodies raised against human CD14 recognize the same doublet of proteins. These results support that mammalian CD14 and the doublet of protein bands in medfly hemocytes share common epitopes. This doublet of protein bands is released from hemocytes upon LPS triggering. A portion of the released protein is clustered on the surface of a distinct hemocyte type and the other remains soluble. The membrane-bound LPS-binding protein is involved in LPS internalization and Escherichia coli phagocytosis but not in LPS signaling.     

The evolutionary dynamics of local infection and global reproduction in host–parasite interactions

A fundamental question in both evolutionary biology and parasitology is why do different levels of virulence evolve in different parasites. Here we use explicitly spatial lattice models to show how the spatial relationships of infection and host reproduction determine the degree of virulence that will occur. When the reproduction of the host acts over larger spatial scales than the infection process higher virulence is predicted. In contrast to both the mean-field and the case where infection acts over larger spatial scales than reproduction, the transmission and virulence predicted are always finite as "self-shading" of infected individuals always occurs. This process may help to explain the evolution of the high virulence of larval diseases of insects where reproduction clearly acts over greater distances than infection.     

Quantification of parasite development in the host-parasite system Biomphalaria glabrata and Schistosoma mansoni

The visceral mass of Biomphalaria glabrata uninfected or infected with Schistosoma mansoni was serially sectioned. The amount of hepatopancreas tissue and of parasite tissue was quantified. In pool-infected snails the volume of the whole visceral mass increased very significantly until week 6 and then decreased. Due to the growing parasites the volume of the visceral mass in infected snails was at most times higher when compared with uninfected snails of the same dimensions. Two weeks p.i. there was a permanent and significant increase of parasite tissue until week 6. During this time the amount of hepatopancreas tissue still increased. From this time onwards till week 12 the proportion of parasite tissue remained rather constant, indicating a kind of regulation, whereas the hepatopancreas tissue decreased to about one-third of the volume found in uninfected snails of the same shell diameter. Compared with infections by a single miracidium there was a significant increase in parasite tissue after infection with two miracidia. Infections with more miracidia (5, 10, and 20) gave no significant further increase. This also demonstrates a kind of regulation. Mortality rate, growth rate and egg production were studied during an infection period of 12 weeks.     

Family-level covariation between parasite resistance and mating system in a hermaphroditic freshwater snail

Genetic compatibility, nonspecific defenses, and environmental effects determine parasite resistance. Host mating system (selfing vs. outcrossing) should be important for parasite resistance because it determines the segregation of alleles at the resistance loci and because inbreeding depression may hamper immune defenses. Individuals of a mixed mating hermaphroditic freshwater snail, Lymnaea ovata, are commonly infected by a digenetic trematode parasite, Echinoparyphium recurvatum. We examined covariation between quantitative resistance to novel parasites and mating system by exposing snail families from four populations that differed by their inbreeding coefficients. We found that resistance was unrelated to inbreeding coefficient of the population, suggesting that the more inbred populations did not carry higher susceptibility load than the less inbred populations. Most of the variation in resistance was expressed among the families within the populations. In the population with the lowest inbreeding coefficient, resistance increased with outcrossing rate of the family, as predicted if selfing had led to inbreeding depression. In the other three populations with higher inbreeding coefficients, resistance was unrelated to outcrossing rate. The results suggest that in populations with higher inbreeding some of the genetic load has been purged, uncoupling the predicted relationship between outcrossing rate and resistance. Snail families also displayed crossing reaction norms for resistance when tested in two environments that presented low and high immune challenge, suggesting that genotype-by-environment interactions are important for parasite resistance.     

Comparative genetic diversity of parasites and their hosts: population structure of an urban cockroach and its haplo-diploid parasite (oxyuroid nematode)

Few studies have investigated the genetic structure of both host and parasite populations at a level of populations and at a level of individuals. We investigated the genetic structure of the urban cockroach Blattella germanica and its oxyuroid parasite Blatticola blattae. Random amplified polymorphic DNA (RAPD) markers were used to quantify genetic diversity between and within four populations (from two cities in France) of the host and its parasite. Diversity based on phenotypic frequencies was calculated for each RAPD marker using Shannon-Wiener's index. We used multivariate analyses to test the significance of genetic differentiation between host and parasite populations. Analysis of molecular variance was also used. Both methods gave similar results. Diversity between pairs of individuals was estimated by Nei & Li's index. Genetic diversity was higher within host or parasite populations (80% and 82%, respectively, of explained diversity) than between host or parasite populations (20% and 18%, respectively, explained diversity). The genetic distances between pairs of parasite populations (or individuals) were not correlated with the genetic distances between the corresponding pairs of host populations (or individuals).     

Paternity-parasitism trade-offs: a model and test of host-parasite cooperation in an avian conspecific brood parasite

Efforts to evaluate the evolutionary and ecological dynamics of conspecific brood parasitism in birds and other animals have focused on the fitness costs of parasitism to hosts and fitness benefits to parasites. However, it has been speculated recently that, in species with biparental care, host males might cooperate with parasitic females by allowing access to the host nest in exchange for copulations. We develop a cost-benefit model to explore the conditions under which such host-parasite cooperation might occur. When the brood parasite does not have a nest of her own, the only benefit to the host male is siring some of the parasitic eggs (quasi-parasitism). Cooperation with the parasite is favored when the ratio of host male paternity of his own eggs relative to his paternity of parasitic eggs exceeds the cost of parasitism. When the brood parasite has a nest of her own, a host male can gain additional, potentially more important benefits by siring the high-value, low-cost eggs laid by the parasite in her own nest. Under these conditions, host males should be even more likely to accept parasitic eggs in return for copulations with the parasitic female. We tested these predictions for American coots (Fulica americana), a species with a high frequency of conspecific brood parasitism. Multilocus DNA profiling indicated that host males did not sire any of the parasitic eggs laid in host nests, nor did they sire eggs laid by the parasite in her own nest. We used field estimates of the model parameters from a four-year study of coots to predict the minimum levels of paternity required for the costs of parasitism to be offset by the benefits of mating with brood parasites. Observed levels of paternity were significantly lower than those predicted under a variety of assumptions, and we reject the hypothesis that host males cooperated with parasitic females. Our model clarifies the specific costs and benefits that influence host-parasite cooperation and, more generally, yields precise predictions about expected levels of host male paternity. These predictions will enable a more rigorous assessment of field studies designed to test adaptive hypotheses of host-parasite cooperation.     

The coevolutionary dynamics of obligate ant social parasite systems--between prudence and antagonism

In this synthesis we apply coevolutionary models to the interactions between socially parasitic ants and their hosts. Obligate social parasite systems are ideal models for coevolution, because the close phylogenetic relationship between these parasites and their hosts results in similar evolutionary potentials, thus making mutual adaptations in a stepwise fashion especially likely to occur. The evolutionary dynamics of host-parasite interactions are influenced by a number of parameters, for example the parasite's transmission mode and rate, the genetic structure of host and parasite populations, the antagonists' migration rates, and the degree of mutual specialisation. For the three types of obligate ant social parasites, queen-tolerant and queen-intolerant inquilines and slavemakers, several of these parameters, and thus the evolutionary trajectory, are likely to differ. Because of the fundamental differences in lifestyle between these social parasite systems, coevolution should further select for different traits in the parasites and their hosts. Queen-tolerant inquilines are true parasites that exert a low selection pressure on their host, because of their rarity and the fact that they do not conduct slave raids to replenish their labour force. Due to their high degree of specialisation and the potential for vertical transmission, coevolutionary theory would predict interactions between these workerless parasites and their hosts to become even more benign over time. Queen-intolerant inquilines that kill the host queen during colony take-over are best described as parasitoids, and their reproductive success is limited by the existing worker force of the invaded host nest. These parasites should therefore evolve strategies to best exploit this fixed resource. Slavemaking ants, by contrast, act as parasites only during colony foundation, while their frequent slave raids follow a predator prey dynamic. They often exploit a number of host species at a given site, and theory predicts that their associations are best described in terms of a highly antagonistic coevolutionary arms race.