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Associated changes in the size and number of mitochondria present in the midgut of the larvae of the housefly, Musca domestica and phospholipid composition of the larvae.

A possible relationship between the phospholipid base composition of larvae of the housefly and the size, number and total volume of midgut mitochondria was examined. The phospholipid composition of the larvae was altered by restricting the amount of choline, substituting choline by N-dimethylethyl-(2-hydroxyethyl)-ammonium salt or 2-N-dimethylaminoethanol and by the addition of 2-aminobutan-1-ol to the larval diet. Mitochondrial measurements were made on electron micrographs of the epithelial cells in the midgut of third instar larvae. Reduction in the normal proportion of phosphatidylcholine in the phospholipids of the larvae was associated with an increase in the average size of the mitochondria and a decrease in their number, probably due to enhanced mitochondrial fusion. The opposite effect was noted when phosphatidyl-2-aminobutan-1-ol was present as a part replacement of the phosphatidyl-ethanolamine normally present in the larvae. Larvae which contained both phosphatidyl-2-aminobutan-1-ol as well as reduced amounts of phosphatidylcholine, contained mitochondria which were intermediate in both size and number. The effects associated with the changes in phosphatidylcholine and phosphatidyl-2-aminobutan-1-ol were independent of each other. The presence of the analogue of phosphatidylcholine and phosphatidyldimethylaminoethanol had no significant effect on the mitochondrial distribution. Total mitochondrial volume remained unaffected by alterations in the phospholipid composition of the larvae.

Animals

Radiolabeling of infective third-stage larvae of Strongyloides stercoralis by feeding [75 Se]selenomethionine-labeled Escherichia coli to first- and second-stage larvae.

A technique is described for radiolabeling Strongyloides stercoralis larvae with [75Se]selenomethionine. Cultures of an auxotrophic methionine-dependent stain of Escherichia coli were grown in a medium containing Dulbecco's modified Eagle's medium supplemented with 5% nutrient broth, amino acids, and [75Se]selenomethionine. When the 75Se-labeled bacterial populations were in the stationary phase of growth, cultures were harvested and the bacteria dispersed on agar plates to serve as food for S. stercoralis larvae. Use of nondividing bacteria is important for successful labeling because the isotope is not diluted by cell division and death of larvae attributable to overgrowth by bacteria is prevented. First-stage S. stercoralis larvae were recovered from feces of infected dogs and reared in humid air at 30 C on agar plates seeded with bacteria. After 7 days, infective third-stage larvae were harvested. The mean specific activity of 6 different batches of larvae ranged from 75 to 330 counts per min/larva with 91.8 +/- 9.5% of the population labeled sufficiently to produce an autoradiographic focus during a practicable, 6-wk period of exposure. Labeled infective larvae penetrated the skin of 10-day-old puppies and migrated to the small intestine, where the developed to adulthood.

Animals

Skin penetration of infective hookworm larvae. II. The path of migration of infective larvae of Ancylostoma braziliense in the metacarpal foot pads of dogs.

The hairless metacarpal foot pads of six hookworm-free puppies were exposed to infective larvae of Ancylostoma braziliense. Serial sections of the biopts stained with Harris' haematoxylin and eosin showed that the infective larvae are able to penetrate the toughest region of canine skin. Pores of eccrine sweat glands did not seem to constitute sites of entry and no larvae were detected in these glands. Larvae were only observed in the epidermis. The histopathology of the infected skin of the foot pads of the puppies was similar to that in human skin with "creeping eruption" as described by Fülleborn (1927). The biopts appeared to consist of hairy skin as well. In the unexposed adjacent hairy skin of the foot pads, larvae were also observed. They were found in the epidermis, hair follicle systems and dermis, suggesting that the migration from the epidermis into deeper tissue depends on the presence of the hair follicle systems.

Ancylostoma

Hyaluronidase from infective Ancylostoma hookworm larvae and its possible function as a virulence factor in tissue invasion and in cutaneous larva migrans.

During skin penetration, infective hookworm larvae encounter hyaluronic acid as they migrate between epidermal keratinocytes and through the ground substance of the dermis. A hyaluronidase would facilitate passage through the epidermis and dermis during larval invasion. Zoonotic hookworm larvae of the genus Ancylostoma were shown to contain a hyaluronidase activity that migrated on modified sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) hyaluronic acid gels with an apparent Mr of 49,000. A second form with an Mr of 87,000 was also identified. The major etiologic agent of cutaneous larva migrans, A. braziliense, was shown to have the greatest enzyme activity, hydrolyzing up to 3.3 micrograms of hyaluronic acid per h per micrograms of total parasite protein at pH 6.0, whereas A. caninum and A. tubaeforme each had much less enzyme activity. The differences in enzyme activities between species correlated with differences in the intensities of the lytic zones at 49 and 87 kDa on SDS-PAGE hyaluronic acid gels. Hookworm hyaluronidase activity exhibited a broad pH optimum between 6.0 and 8.0 and did not hydrolyze chondroitin sulfate, two features that suggest that the hookworm enzyme is more like the invertebrate leech hyaluronidase than mammalian testicular or lysosomal hyaluronidase. Larvae of A. braziliense were shown to release hyaluronidase activity and degrade radiolabeled hyaluronic acid in vitro. Gold sodium thiomalate was identified as an enzyme inhibitor. The hyaluronidase is the second major virulence factor that we have identified from infective hookworm larvae.

Ancylostoma

Skin penetration of infective hookworm larvae. I. The path of migration of infective larvae of Ancylostoma braziliense in canine skin.

Migratory behaviour of Ancylostoma braziliense was studied in relation to the structure of the skin in dogs after primary infections. Data were obtained studying serial sections of lateral skin areas 6 mm in diameter, which had been exposed to larvae. The sections were stained either with Harris' haematoxylin and eosin or with P.A.S. or as outlined by Crossmon. Most of the larvae managed to penetrate the skin within 1/2 hr after the application. Hairs did not seem to constitute sites of entry. The larvae moved into the horny layer where edges of keratinized cells provide uneven spots. They migrated approximately parallel to the surface from the horny layer into the living epidermis and continued into an external root sheath of a hair follicle. They could only leave this site via sebaceous glands for the dermis or via apocrine sweat glands for the hypodermis. Tunnels from the epidermis into the dermis, however, suggested that a direct trans-epidermal migration had occurred. The vessels invaded by larvae were hypodermal lymphatic vessels. The first ones were found in these structures 1/2 h after the onset of the exposure.

Ancylostoma

The free swimming Pipa larvae, with a review of pipid larvae and pipid phylogeny (Anura: Pipidae).

This paper describes the morphology of the free swimming Pipa larvae, compares them with Xenopus, Hymenochirus, and to some extent, Rhinophrynus larvae, and presents a morphological diagnosis of pipid larvae. Pipa and Xenopus have very similar chondrocrania. Hymenochirus is superficially different but has the same diagnostic features. The differences appear related to its small size and predatory habitus. Other aspects of anatomy, especially the filter apparatus are very different in each genus. The filter apparatus of Pipa is somewhat reduced and seems modified for the retention of relatively large (20+ microns) particles. Similar adaptations may have been annectant to predations in Hymenochirus, which lacks a filter apparatus. However, varying states of seven character complexes, which cut across the varying ecology, show that there are two basic pipid lineages, each currently confined to Africa or South America, respectively. Recent finds of fossil South American Xenopus indicate that these two lineages separated before the continents did. This does not warrant the recognition of two subfamilies because Xenopus and Hymenochirus are too different. Pseudhymenochirus is not an intermediate between them; it is primitive Hymenochirus. Eight character states separate pipid and rhynophrynid larvae.

Animals

Scanning electron microscopy of the sheathed infective larva and parasitic third-stage larva of Haemonchus contortus (Nematoda: Trichostrongyloidea).

Scanning electron microscopy was used to describe the infective and parasitic third-stage larvae of Haemonchus contortus, the large stomach worm of ruminants. Infective larvae are ensheathed in the cuticle of the second stage, so the descriptions are of the second- and third-stage cuticles. Both larval stages had an inner circle of 6 labial papillae, an outer circle of 6 labial papillae and 4 somatic papillae, and lateral amphidial pits. Infective larvae (cuticle of the second stage) had the 6 internal labial papillae on prominent bluntly rectangular lappets in a star-shaped arrangement around a large triradiate mouth, small triangular or round amphidial pits, flattened ribbonlike lateral alae, and phasmidial apertures opening on the ventral surface of the lateral alae. Parasitic third-stage larvae had the 6 internal labial papillae on small elevations without lappets around a small mouth; large, oval amphidial pits; ribbonlike lateral alae for most of their length, but with the anterior 30-40 microns of the alae cordlike; and phasmidial apertures on the body cuticle ventral to the lateral alae.

Animals

Trichinella spiralis: activation of complement by infective larvae, adults, and newborn larvae.

The ability of Trichinella spiralis to activate complement (C) has been addressed by several investigators. However, these investigators employed methods in which either detection of C fragments on the parasite surface or the adherence of leukocytes to the parasite was considered an indication of C activation. The present studies were undertaken to examine: (a) whether activation of C occurs via the classical and/or alternative pathway, (b) at which stage(s) of the parasite C activating capacity is acquired, and (c) what molecular entities of the epicuticle and/or cuticle are responsible for initiating C activation. Our studies indicate that T. spiralis activates C primarily via the alternative pathway (and weakly via the classical pathway) since incubation of parasites obtained from infected mice with either normal human serum (NHS) or Mg.EGTA-NHS, followed by incubation (1 hr, 37 degrees C) with antibody-sensitized sheep erythrocytes or rabbit erythrocytes, respectively, showed a time-and parasite number-dependent depletion of C. Although the three stages of T. spiralis, i.e., infective larvae, adults and newborn larvae, are capable of activating C, the newborn appears to be the most potent activator, especially when parasite number and size are taken into consideration. Further evidence of C activation is obtained from SDS-PAGE and Western blot analysis in which homogenates of parasites preincubated with NHS showed the presence of C3, C9, and C1q, whereas controls without serum were negative. Since isolated C1q was also capable of directly binding to the surface of adults and infective larvae, it is postulated that their cuticle and/or epicuticle may possess surface structures which serve as binding sites for C1q.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

The effects of time and temperature of storage on the infectivity of third-stage larvae of Heligmosomoides polygyrus (= Nematospiroides dubius). 2. Studies on the fecundity of female worms as a function of the infectivity of the third-stage larvae.

In this study we examined the effect of infectivity of third-stage larvae on the fecundity of H. polygyrus female worms. Groups of 10 mice were infected with third-stage larvae which had been stored for different lengths of time at 22 degrees C. Results showed that the number of eggs laid by one female worm increased when the worm burden was smaller and decreased with a greater worm burden. Also, the egg-output of female worms growing in a similar population varied according to the level of infectivity of the larvae, with a low infectivity resulting in greater fecundity and vice versa. These results may explain the variations in egg-output which are recorded at different times of the year from the parasites of grazing animals. If this phenomenon is confirmed in other species of parasites, it will be important for epidemiological studies.

Animals

Larva-specific antibodies in patients with visceral larva migrans.

Seven of 10 patients with visceral larva migrans (VLM) had serum precipitating antibodies specific for larval antigens of Toxocara canis as determined by double diffusion in agar. Such antibodies were absent in 114 normal adults and 25 patients with collagen disease. Precipitation of ascarid components by C-reactive protein resulted in false-positive reactions, but this precipitation was readily prevented by appropriate absorption of sera before testing. A more sensitive assay, the enzyme-linked immunosorbent assay, revealed high titers of antibodies to larvae of Toxocara in all patients with VLM; the log2 titer ranged from 9 to 14. Five of eight household relatives of these patients and four children who had had VLM two to four years before testing had titers of 6-12. Of the 114 normal adults, 105 had titers of 0-2; nine had titers of 3-8. Of the 25 patients with collagen disease, 23 had titers of 0, and two had titers of 4 and 6, respectively. Additional studies of those patients indicated that infection with Toxocara can be distinguished serologically from ascariasis, filariasis, and trichinellosis.

Antibodies

A comparison of the proteins found in developing wild type larvae and developing lethal mutant larvae of Drosophila melanogaster.

Extracts of late larval lethal mutants were compared with extracts of wild type larvae of the same developmental age on double diffusion plates using 16 different antisera. Nearly all of the mutant extracts showed relative antigen concentration differences compared with the wild type and four of the mutants lacked a protein at death found in the wild type of the same developmental age. In each case it was a different protein. The results are discussed by considering the different ways in which mutations can lead to the loss of a protein in developing systems.

Animals