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N A Ratcliffe

Publications and source records attributed to N A Ratcliffe.

At least 55 records · Page 3Linked to original sources

Comparative study of hemocytes and associated cells of some medically important dipterans.

The aim of this work is to study, characterize, and compare different morphological types of hemocytes of Glossina austeni, G. morsitans, Calliphora erythrocephala, Stomoxys calcitrans, Lucilia sericata, Aedes aegypti, and Culex quinquefasciatus. This information is intended to provide a basis for future studies of the cellular defense mechanisms of these dipterans. Seven morphological types of hemocytes were identified by phase-contrast optics: prohemocytes, plasmatocytes, thrombocytoids, granulocytes, adipohemocytes, oenocytoids, and spindle cells of various sizes. Adipohemocytes are difficult to distinguish from both fat body cells and granulocytes. All seven cell types are not present in every species. For example, thrombocytoids and spindle cells were not found in A. aegypti or C. quinquefasciatus, and oenocytoids were observed only in A. aegypti, C. quinquefasciatus, and in C. erythrocephala. In addition to the hemocytes, fat body cells and nephrocytes are also freely present in the hemolymph of some species but may have gained access to the blood during the bleeding process. With electron microscopy and with thick plastic sections of G. austeni hemolymph, only nephrocytes, plasmatocytes, granulocytes, and spindle cells were identified with any certainty, and the large spindle cells are morphologically very different from those found in the other dipterans. They are rigid cells supported by microtubules running throughout their entire length. These hemocytes are present in large numbers only in newly emerged flies, they are absent in larvae and young pupae, and are rare in old adults. Their disappearance from the hemolymph of newly emerged Glossina appears to be a result of phagocytosis by the plasmatocytes.

Anatomy, Comparative↗

Evidence for the presence of subpopulations of Arenicola marina coelomocytes identified by their selective response towards Gram+ve and Gram-ve bacteria.

In Arenicola marina (L.), in vitro attachment and phagocytosis of bacteria appears to be serum-independent and thus, non-specific. Mixtures of Gram+ve and Gram-ve bacterial species (Gram+ve/-ve, of two Gram+ve (Gram+ve/+ve) and of two Gram-ve bacterial species (Gram-ve/-ve)were overlaid on coelomocyte monolayers, in order to introduce competition for cell surface 'receptors'. An enhanced total phagocytosis was only recorded in the Gram+ve/-ve mixtures in comparison with their controls. The results are interpreted in terms of coelomocyte subpopulations and indicate that A. marina coelomocytes may possess specific carbohydrate/glycoprotein determinants capable of recognizing basic carbohydrate differences between the cell walls of Gram+ve and Gram-ve bacteria.

Alcaligenes↗

Integumental grafting and immunorecognition in insects.

In preliminary experiments, integumental grafting was conducted on a range of insect species in order to elucidate levels of insect immunorecognition. Subsequently, however, only Blaberus craniifer and Extatosoma tiaratum were utilised for detailed study because of their high survival rates and longevity. In both host species, most autografts and allografts rapidly healed and were accepted while xenografts usually became melanized and were rejected. A significant correlation was obtained between the mean rejection times, which ranged from 8.0 +/- 1.8 days to 28.2 +/- 6.6 days, and the phylogenetic relatedness of the xenograft donors to the host species. A preliminary histological investigation revealed the accumulation of blood cells below the grafts, a process that was much more marked with xenografts than with autografts.

Animals↗

Insect erythrocyte agglutinins. In vitro opsonization experiments with Clitumnus extradentatus and Periplaneta americana haemocytes.

The effect of naturally occurring haemagglutinins on the in vitro phagocytosis of sheep erythrocytes by the blood cells (haemocytes) of Clitumnus extradentatus and Periplaneta americana was studied. The results showed that the haemagglutinins in both species failed to act as opsonins. Indeed, in some instances, incubation of erythrocytes in haemolymph resulted in less avid ingestion as compared with the saline-incubated controls. This reduced phagocytosis was probably caused by the clumping of erythrocytes on the haemocyte monolayers, leaving fewer single red cells available for uptake. The possible roles of these erythrocyte agglutinins in the host defence systems of insects are discussed.

Animals↗

An ultrastructural and cytochemical study of the interaction between latex particles and the haemocytes of the wax moth Galleria mellonella in vitro.

The plasmatocytes are the major phagocytic blood-cell type in the haemolymph of the wax-moth, Galleria mellonella. In the present study, these cells were allowed to attach to tissue culture dishes for 1 h, rinsed and then incubated with latex beads for up to 72 h. These cells were then fixed for routine transmission electron microscopy and acid phosphatase cytochemistry. Intracellular latex particles were found in tight, ill-defined phagosomes, which were often clearly associated with the Golgi complexes of the plasmatocytes. Fusion of both primary lysosomes and multivesicular bodies with the phagosomes occasionally occurred and this resulted in the accumulation of an acid phosphatase positive reaction product around the test particles. Subsequent experiments showed that this acid phosphatase activity was mainly associated with the primary lysosomes. The results of the lysosome/latex interactions are compared with those obtained from similar studies on the digestive mechanisms in other phagocytes.

Acid Phosphatase↗

Studies on the in vivo cellular reactions of insects: an ultrastructural analysis of nodule formation in Galleria mellonella.

Cellular aggregates or nodules which formed in Galleria mellonella larvae in response to injections of killed Bacillus cereus were examined during their first 24 hr of development. The initial aggregation stage was very rapid, occurring within 1 min, and was brought about by the release of an adhesive flocculent material by granular haemocytes in contact with the bacteria. This material, which subsequently became increasingly electron dense owing to melanin deposition, consolidated and by 5 min ramified all the intercellular spaces. The granular cells degenerated, releasing portions of cytoplasm form their peripheries, and eventurally appeared in the matrix as voids containing loosely flocculent material. No bacterial breakdown was seen, however. From 2-4 hr, plasmatocytes began to encapsulate the necrotic mass, and by 24 hr a complete sheath, composed of three discrete regions of cells, was formed. These findings are discussed in relation to the various proposed mechanisms of nodule and capsule formation.

Animals↗

The granular cells of Galleria mellonella during clotting and phagocytic reactions in vitro.

Light and electron-microscopic observations of the blood-cells (hemocytes) of the wax-moth Galleria mellonella showed that hemolymph coagulation was initiated by the rapid release of material from the granular cells. During incubation for short terms in vitro these cells showed progressive degranulation as material derived from the granules was discharged into the hemolymph. Attempts to determine the nature of this material by staining with ruthenium red proved mainly unsuccessful. When challenged with bacteria in vitro the granular cells failed to phagocytose these particles and instead the bacteria became embedded in the granular material surrounding these cells. The mode of coagulation reported here is compared with previous reports of the role of invertebrate hemocytes in hemolymph clotting.

Animals↗