Immunoparasitology in Avian Species

There has been no work on the immunological response of birdsto helminth infections since the late 1960s, an area of investigationthat has been too long ignored. Similarly, studies of arthropod-mediatedresponses in birds are lacking except for a few scattered investigations.Recently, a serum antibody response has been seen against onearthropod, the northern fowl mite. The appearance of antibodiesrecognizing an 8–10 kilodalton mite antigen seems to correlatewith a reduction in the mite population on infested chickens.Most of the studies on parasite immunity in avian species havecentered on the economically important Eimeria species, protozoanparasites that infect the intestine of chickens and turkeys.These investigations encompass wide areas of interest includingthe effect of immunity on parasite invasion, development ofT-cell proliferation assays and T-cell clones, inhibition ofparasite penetration and development by hybridoma antibody treatment,production of genetically engineered Eimeria antigens used inbird immunization studies, and studies using inbred or congeniclines of birds to determine what effect the major histocompatibilitycomplex has on parasite immunity. From these efforts it hasbeen learned that not only is the immunity species-specific,but also depending on where in the intestine the parasite invades,penetration is either not affected or inhibited by as much as50%. The T-cell proliferation assays suggest that this specificitymay be due to a species-specific T-cell response. Immunizationstudies using a genetically engineered antigen have indicatedthat at least partial protection against one species of Eimeriais possible. Studies done with the inbred congenic lines ofbirds have shown that the genetic makeup of the bird is importantin how it responds to either a natural infection or to immunizationwith a genetically engineered antigen. Clearly, these resultsshow not only the complexity of the bird response to parasiteinfection, but also the amount of work still undone.     

The evolution of hymenopteran wings: the importance of size

The allometric relationships between body size and several aspects of wing morphology in the insect order Hymenoptera were investigated using multivariate morphometric techniques. The study focused primarily on wing allometry in five monophyletic genera of bees ( Perdila, Halictus, Ceratina, Trigona and Apis ), but the patterns of size-related evolutionary change found within each of these genera are also found to exist in numerous other hymenopteran taxa. Increased body size in hymenopteran lineages is correlated with the following changes in wing morphology: (1) decreased relative stigma area, (2) distal extension of wing vein elements, (3) increased aspect ratio and (4) proximal shift in the centroid of wing area. The reverse is true for decreased size. The widespread allometric trends most likely result from adaptive change in wing morphology due to size-related changes in the physical properties impinging on the organism–principally the quality and magnitude of drag. The fact that similar wing morphologies among distantly related species can result from similarity in body size has important implications for the study of hymenopteran phylogeny, especially at lower taxonomic levels and when a high proportion of wing characters are employed.     

Identification of an Apically-Located Antigen That Is Conserved in Sporozoan Parasites

ABSTRACT. Sporozoan parasites of the phylum Apicomplexa all possess common apical structures. The current study used a monoclonal antibody (mAb-E12) to identify a conserved antigen in the apical region of merozoites of seven species of Plasmodium (including rodent, primate and human pathogens), tachyzoites of Toxoplasma gondii , bradyzoites of Sarcocystis bovis , and sporozoites and merozoites of Eimeria tenella and E. acervulina. The antigen was also present in sporozoites of haemosporinid parasites. Immunofluorescence studies showed that the antigen was restricted to the apical 3rd of these invasive stages. Using immunoelectron microscopy, labeling was demonstrated in the region of the polar ring, below the paired inner membranes of the parasite pellicle, and near the subpellicular microtubules radiating from the polar ring of merozoites and sporozoites of E. tenella . The majority of the antigen could be extracted with 1% Triton-X 100, but a portion remained associated with the cytoskeletal elements. The molecule has a relative rate of migration (M_r) of 47,000 in Plasmodium spp. and 43–46,000 in coccidian species. Since the epitope recognized by mAb-El 2 is highly conserved, restricted to motile stages, and appears to be associated with microtubules, this antigen could be involved in cellular motility and cellular invasion.     

Effects of Cationized Ferritin and Neuraminidase on Invasion of Cultured Cells by Eimeria meleagrimitis Sporozoites

Primary turkey kidney cells and Eimeria meleagrimitis sporozoites were treated with cationized ferritin (CF) or neuraminidase (NANase), and the effects on the invasion of the cells by the sporozoites were measured. Cultures of host cells pretreated with either compound contained significantly fewer intracellular sporozoites than did control cultures. There was little additive effect if cultures were first treated with NANase and then with CF. In contrast, pretreatment of sporozoites with CF or low concentrations of NANase had no effect on invasion. The inhibition of invasion was apparently due to an interaction between treatment substances and host cell surface rather than to direct effect on the sporozoites. The CF bound to the randomly distributed anionic sites on the surfaces of both host cells and sporozoites and then rapidly aggregated. Sporozoites, probably in the process of invading cells, were invariably found with the conoid in close association with aggregates of CF on the host cell membrane. The CF on the sporozoites was apparently shed before or during invasion because all intracellular sporozoites were completely devoid of the label.     

Surface Changes Induced by Immune Serum on Eimeria tenella Sporozoites and Merozoites

ABSTRACT. The surface of merozoites and sporozoites of Eimeria tenella was affected by incubation with E. tenella -immune chicken serum (ICS). Normal chicken serum (NCS) and heat-inactivated ICS had no effect on the pellicular surface of either developmental stage. Sporozoites formed surface bulges or swellings after 10 min of incubation with ICS, and by 15 min postincubation, the morphology of the sporozoites was distorted by a surface coating of fibrinous material. Merozoites exposed to ICS were similarly coated, but surface swelling was not as severe. The coating formed rapidly and was seen as early as 5 min postincubation. Sporozoites incubated with heat-inactivated ICS supplemented with normal chicken serum were coated with a fibrinous material and in some cases lysed. These data indicated that complement must be present for the surface interaction to occur.     

Phylogeny of the bee family Melittidae (Hymenoptera: Anthophila) based on combined molecular and morphological data

The bee family Melittidae comprises a small, but biologically fascinating, group of mostly oligolectic bees, some of which are oil collecting. Phylogenetic relationships within this family are poorly understood and some genera cannot be placed with confidence at the subfamily level. We analysed melittid phylogeny using a combined dataset of five nuclear genes [28S, elongation factor‐1α (EF‐1α, F2 copy), long‐wavelength rhodopsin, Na‐K ATPase and RNA polymerase II] spanning 4842 bp plus 68 adult morphological characters. Our study included 25% of the species‐level diversity and 81% of the generic‐level diversity and included all previously recognized tribes and subfamilies. We analysed the dataset using parsimony, maximum likelihood and Bayesian methods. All methods yielded congruent results. All topologies recovered the three previously recognized subfamilies (Dasypodainae, Melittinae, Meganomiinae), but two genera (Afrodasypoda and Promelitta) are transferred from Dasypodainae to Melittinae. On the basis of our tree topologies we identify four tribes (Dasypodaini comb.n. , Hesperapini stat.n. , Macropidini comb.n. and Melittini), only one of which (Melittini) matches a widely used classification. Lastly, we discuss the evolution of host‐plant association in the light of our new phylogenetic hypothesis. Our results strongly support multiple independent origins of oil‐collecting behaviour in the Melittinae.     

Sporozoites of Mammalian Malaria: Attachment To, Interiorization and Fate Within Macrophages

Sporozoites of Plasmodium berghei and Plasmodium knowlesi, incubated in normal serum readily interact with peritoneal macrophages of mice or rhesus monkeys, respectively. Interiorization of the sporozoite requires that both serum and macrophages be obtained from an animal susceptible to infection by the malaria parasite. Serum requirements for sporozoite attachment to the macrophage are less specific. Phagocytosis is not essential for the parasites to become intracellular. Our findings indicate that active penetration of the sporozoites into the macrophages does occur.     

The Relation of Phosphate Metabolism to Oxidative Assimilation of Acetate by Euglena gracilis var. bacillaris. Effects of 2,4-Dinitrophenol*

SYNOPSIS. Oxidative assimilation of acetate by Euglena gracilis var. bacillaris occurs without appreciable net transfer of inorganic phosphate between cells and medium. Low concentrations of the “uncoupling agent” 2,4-dinitrophenol (2–5 μM, pH 5) stimulate acetate oxidation and inhibit acetate assimilation. These dinitrophenol concentrations have no measurable effect on edogenous respiration. Higher concentrations inhibit respiration on acetate, and still higher concentrations inhibit endogenous respiration. Dinitrophenol concentrations which stimulate acetate oxidation produce no measurable change in the total ATP content of the euglena cells. Higher concentrations (50–100 μM) cause progressive decrease in the ATP content. A hypothetical mechanism is proposed which accounts for the normal, constant oxidation-assimilation ratio for acetate as the result of coupling of oxidative ATP synthesis with assimilatory ATP consumption. The data are consistent with this mechanism if it is additionally assumed that oxidation and assimilation of acetate occur in an intracellular “compartment” separated from other sources and sinks of ATP.