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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

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

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

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

Eosinophils versus neutrophils in host defense. Killing of newborn larvae of Trichinella spiralis by human granulocytes in vitro.

Eosinophil leukocytes have been reported to have a major role in host defense against invasive, migratory phases of helminth infestations, yet the relative larvicidal abilities of eosinophils and neutrophils have not been thoroughly examined. This study examined the killing of newborn (migratory phase) larvae of Trichinella spiralis during incubation by human granulocytes in vitro. The assay employed cultue of larvae with cells, sera, and reagents in microtiter wells with direct counting of surviving larvae after incubation. Killed larvae appeared to be lysed. Verification of the microplate assay was obtained by demonstrating complete loss of infectivity of larvae incubated with leukocytes and immune serum. In the presence of optimal immune serum concentrations, purified neutrophils or eosinophils achieved >/=95% killing of larvae at cell:larva ratios of 2,000:1 or greater. Fresh normal serum prompted slight (19%) killing by leukocytes at a cell:larva ratio of 9,000:1. Cells plus heat-inactivated normal serum and all sera preparations in the absence of leukocytes killed <8% of the larvae. The activity of immune serum was opsonic. Cells adhered to larvae that had been preincubated in immune serum, and immunofluorescent studies indicated that such preopsonized larvae were coated with immunoglobulin (Ig)G. However, preopsonized larvae lost opsonic activity and surface IgG during incubation for 3 h in medium lacking immune serum. The rate of killing was dependent on the cell:larva ratio; at high leukocyte concentrations (4,200:1), 99% were killed within 7 h; at lower cell:larva ratios, killing increased steadily during a 20-h incubation period. Killing was inhibited by 20 mug catalase, 5 mug/ml cytochalasin B, or 5muM colchicine, but was unchanged by superoxide dismutase and was enhanced by azide or cyanide. Leukocytes from a patient with chronic granulomatous disease, lacking ability to mount a normal oxidative response, demonstrated a markedly suppressed larvicidal effect. The data indicate that neutrophils are at least as effective as eosinophils in the killing of newborn larvae of T. spiralis. The killing appeared to be mediated by the oxidative metabolic burst with its generation of hydrogen peroxide.

Animals

Boophilus microplus: rejection of larvae from British breed cattle.

Larvae of Boophilus microplus were labelled with [32P] and used to study the rejection of larvae from British breed cattle with different levels of resistance. On animals of high resistance the loss of larvae due to grooming ranged from 9-54% during the first 24 h of infestation, and more time was spent grooming by these animals. Most of the grooming activity could be attributed to the presence of tick larvae and the presence of older tick stages did not increase the percentage of larvae lost. Grooming was directed to attached larvae and these could be removed. Animals of low resistance did not lose a significant number of larvae as a result of grooming, but all previously infested hosts lost a proportion of the larvae (18-39%) which could not be accounted for by grooming. This loss was greater than the total loss of larvae (11%) on animals not previously exposed to B. microplus.

Animals

Emerging trends in the study of spiralian larvae.

Many animals undergo indirect development, where their embryogenesis produces an intermediate life stage, or larva, that is often free-living and later metamorphoses into an adult. As their adult counterparts, larvae can have unique and diverse morphologies and occupy various ecological niches. Given their broad phylogenetic distribution, larvae have been central to hypotheses about animal evolution. However, the evolution of these intermediate forms and the developmental mechanisms diversifying animal life cycles are still debated. This review focuses on Spiralia, a large and diverse clade of bilaterally symmetrical animals with a fascinating array of larval forms, most notably the archetypical trochophore larva. We explore how classic research and modern advances have improved our understanding of spiralian larvae, their development, and evolution. Specifically, we examine three morphological features of spiralian larvae: the anterior neural system, the ciliary bands, and the posterior hyposphere. The combination of molecular and developmental evidence with modern high-throughput techniques, such as comparative genomics, single-cell transcriptomics, and epigenomics, is a promising strategy that will lead to new testable hypotheses about the mechanisms behind the evolution of larvae and life cycles in Spiralia and animals in general. We predict that the increasing number of available genomes for Spiralia and the optimization of genome-wide and single-cell approaches will unlock the study of many emerging spiralian taxa, transforming our views of the evolution of this animal group and their larvae.

Animals

Infectivity of Ancylostoma caninum larvae after freezing over liquid nitrogen.

Third-stage larvae of Ancyclostoma caninum were exsheathed either by carbon dioxide or sodium hypochlorite treatment. Exsheathment per se reduced the infectivity of third-stage larvae by 45% to 59% without affecting larval motility. Both ensheathed and exsheathed larvae were frozen over liquid nitrogen for 90 days, and thawed. Frozen ensheathed larvae were all killed; but frozen exsheathed larvae were 40-54% as infective as non-frozen exsheathed larvae, and the addition of 10% dimethyl sulphoxide raised the infectivity to 89%, relative to non-frozen exsheathed larvae. Taken together, the deleterious effects of exsheathment and freezing resulted in an infectivity of 16-37%, relative to normal larvae.

Ancylostoma

Observations on the infectivity of parasitic third-stage larvae of Uncinaria lucasi Stiles 1901 (Nematoda: Ancylostomatidae) of Northern fur seals, Callorhinus ursinus Linn., on St. Paul Island, Alaska.

Twelve fur seal pups, which had not nursed their mothers, were used in an infectivity experiment. Pups were exposed to parasitic 3rd-stage larvae of Uncinaria lucasi from belly tissues of fur seal bulls, bachelors, and pregnant cows, to determine maturation capability of the larvae. Hookworms were not recovered from the intestines of 3 pups receiving larvae from belly blubber of bulls, 6 pups receiving larvae from belly blubber of bachelors, and 1 nonexposed pup. Maturation of hookworms did occur in 2 pups exposed to larvae from a mixture of belly blubber, mammary tissue, and milk of pregnant cows. Parasitic 3rd-stage hookworm larvae from belly tissues of pregnant and "non-pregnant" fur seal cows averaged 938.1 and 802.1 micron long, and 34.1 and 31.5 micron wide, respectively; however, larvae from belly tissues of a fur seal bull, bachelors, 2-year-old males, male and female yearlings and pups, and Steller Sea Lion subadults averaged 640.5-732.0 micron long and 20.9-24.9 micron wide.

Abdominal Muscles

[Method of selection of synchronously developing larvae of Drosophila melanogaster].

Methods of obtaining synchronously developing larvae were comparatively evaluated. Larvae were selected by distinct morphological features: time of hatching, second larval moult, pupation. Both virgin and non-virgin females were taken in the experiment. The degree of synchronism was estimated by the interval during which pupation took place in all larvae of the sample. When selecting larvae at the time of second moult, the pupation of all selected larvae was completed within 8 hrs. All other methods of obtaining synchronously developing larvae give a spread of pupation from 12 to 25 hrs. A simple modification of the method of larval selection during the second moult is proposed which allows to obtain a large amount of synchronously developing larvae of the 3rd instar.

Animals

The bionomics of the free-living larvae and the transmission of Dictyocaulus filaria between lambs in North-East England.

The bionomics of the free-living larvae of Dictyocaulus filaria on pasture, and the transmission of infection between lambs, were studied during different seasons of the year in North-East England. The rate of development of first-stage larvae to the third stage took 4-9 days in late spring and summer, 1 1/2-4 weeks in autumn and 5 1/2-7 weeks in winter. The proportion of first-stage larvae developing to the third stage ranged from 10-28% in autumn and winter, and 2-25% in spring and summer. The rate of mortality of the third-stage larvae was approximately logarithmic in nature, although the survival time was shorter in spring and summer than in autumn and winter. Third stage larvae were able to survive from autumn until the spring of the following year in sufficient numbers to perpetuate transmission but not to cause clinical disease. In a transmission study, the survival of the infective larvae on the experimental plot was poor in summer, but the larval population increased in the autumn and then declined slowly throughout the winter. Infection in the susceptible lambs was related to the level of infection on the herbage increasing in severity from early summer to late autumn. However, those lambs infected in the summer were resistant to the heavy autumn challenge of larvae on pasture.

Animals