Metabolic differentiation of bloodstream forms of Trypanosoma brucei brucei into procyclic forms. Effect of hydroxyurea, arabinosyl adenine, and serum omission

Bloodforms of Trypanosoma brucei brucei STIB 247 taken from rats and containing more than 80 per cent short stumpy forms, differentiated in vitro to procyclic forms in medium SDM 79 (Brun and Sch?nenberger 1979), enriched with 3 mmol.dm-3 cis-aconitate. Cell division was abolished by the addition of hydroxyurea (200 micrograms.ml-1) or arabinosyl adenine (20 micrograms.ml-1 to the cultivation medium, or by the omission of serum from the medium. The ultrastructure of exponentially growing controls was rearranged within 24 h. The endogenous respiration and the respiration stimulated by proline, succinate, and 2-oxoglutarate were detectable within 12 h; after 48 h the respiration rates were comparable to those found in the established procyclic forms. After 12 h the respiration was inhibited by 200 mumol.dm-3 KCN, and by 20 mumol.dm-3 antimycin to the extent found in procyclic forms. Hydroxyurea did not significantly affect respiration. Activities of procyclic-stage enzyme markers malate dehydrogenase, threonine dehydrogenase, succinate: cytochrome c reductase, and NADH: cytochrome c reductase rose within 48 h of differentiation to values which were close to those found in established procyclic forms. The activity of glutamate dehydrogenase (NAD-specific), however, was only 1/3 of that in the procyclics, and no citrate synthase was detected in differentiating culture. Glycosomal malate dehydrogenase was detected after 6 h. In the presence of hydroxyurea or arabinosyl adenine, or in the absence of serum, respiration rates, marker enzyme activities, and glycosomal malate dehydrogenase developed to the extent comparable to the untreated controls. The results suggest that it is possible to separate the process of differentiation from cell proliferation. Cell division is not a necessary prerequisite of differentiation.     

Ultrastructure of the excretory system of Brachylaimus aequans (Trematoda: Brachylaimoidea)

The ultrastructure of the flame cell, excretory capillaries, ducts, collecting ducts, excretory bladder, and excretory pore of Brachylaimus aequans was studied 6-8 days p.i. The excretory ducts, collecting ducts and excretory bladder are provided with numerous lamellae on the luminal side. The cilia of lateral flames in the excretory ducts have a triated rootlet. The excretory pore is covered by a tegument identical with the body tegument. The syncytium of the excretory bladder is connected with the tegument of the excretory pore by means of a septate desmosome. No lipid or excretory corpuscles have been demonstrated in the excretory system.     

Colonization of Ascaris lumbricoides eggs by the fungus Verticillium chlamydosporium Goddard.

The process of colonization of Ascaris lumbricoides eggs by the fungus Verticillium chlamydosporium was studied by scanning electron microscopy. The preparations were made by fractionation of egg suspension exposed to the fungus for four days and frozen in liquid nitrogen according to St?rba and Milácek (1986). Ovicidal fungus forms an abundant ramifying mycelial network in the area between the eggs. However, egg-shells are penetrated only by some hyphae without any penetration organs produced (simple hyphal penetration). In a liquid medium, after penetration, hyphae inside the eggs rapidly grow among inner structures of egg-shells and on the surface of developing larvae. In the next phase, hyphae colonize the developing larva. The eggs attacked by this fungus remain morphologically unchanged for a long time except the sites of penetration. Verticillium chlamydosporium is a fungus with unique ovicidal properties. It colonizes the eggs of Ascaris lumbricoides at all stages of embryo development and also attacks larvae inside the eggs.     

Investigation of sandflies (Diptera, Phlebotomidae) in an endemic focus of visceral leishmaniasis in Yugoslavia

The paper presents the results of faunistic, ecological and viral investigations concerning phlebotomine sandflies in an endemic focus of visceral leishmaniasis in Yugoslavia. These investigations were carried out in the period from 1969 to 1981.     

Ultrastructure of a new type of sensory ending in Echinostoma revolutum cercaria (Trematoda: Echinostomatidae)

Two pairs of hitherto unknown multiciliate sensory endings have been demonstrated in free-swimming cercariae of Echinostoma revolutum using transmission and scanning electron microscopy. This type of receptor consists of a bundle of 36--40 short, conically projecting cilia ensheathed by a collar formed by a circular evagination of tegument in shape of a cup. The cilia projecting from the centre of a widened nerve fibre, filled with electron-lucid vesicles, possess a well developed basal body and do not contain a ciliary rootlet. The function of this receptor is discussed.     

Cono- and seminiphagous insects of Abies alba Mill. in the Gorce National Park in Poland during 1996?–?1998

A study was conducted on the insects infesting seeds and cones of Abies alba Mill. in the Gorce National Park, in southern Poland during 1996–1998. A total of 856 cones were collected from 66 trees growing at 11 sites located in partially or strictly protected forest reserves. Of this sample, 217 cones with 54,958 seeds were dissected. This study yielded 5,816 specimens of insects, namely: seminiphagous Resseliella piceae Seitn., Megastigmus suspectus Borr. as well as conophagous Earomyia impossibile Morge, Barbara herrichiana Obr. and Dioryctria abietella (Den. et Schiff.). The most abundant species was R. piceae with 4,186 individuals. The index of cone infestation was calculated. It showed great variability between forest reserves and study-years. The numbers of insect individuals, numbers of viable and infertile seeds, and seeds damaged by M. suspectus and R. piceae, in both strict and partial reserves in the consecutive years were compared statistically. The test of significant differences between two frequencies at 0.05 level was used in statistical analysis. In both strict and partial reserves, statistically significant differences were found between studied variables.     

Studies on insects and mites causing galls on the leaves of purple willow Salix purpurea L. in the Ojców National Park in Poland

Analytical studies were conducted on the species composition of galls occurring on the leaves of Salix purpurea L. in the Ojców National Park in 1999–2000. The species composition of galls was identical in two consecutive years. Galls were caused by 5species (a total of 480galls). The most numerous species in both years was Pontania viminalis (L.). Index of occurrence frequency and Agrell's index of species co-existence have been calculated. This index reached the highest values in case of Pontania dolichura (Thoms.) and P. viminalis (L.) in 1999 as well as Phyllocolpa leucaspis (Tischb.) and P. viminalis (L.) in 2000.     

An in vitro study of hematopoietic progenitor cells from peripheral blood of rabbits

The purpose of this study was to isolate and cultivate a subpopulation of pluripotent stem cells (PSCs) from the peripheral blood of rabbits, which are frequently used in veterinary research as an animal model. Pluripotent stem cells, as described in human beings, are fibroblast-like cells that exhibit a CD34 marker, specific from other hematopoietic stem cells. Commonly used human commercial media has been researched for culturing rabbit PSCs. These findings allow us to contemplate the direct application of this simple and standardized methodology in several areas of study, such as of the pharmacological effect of many drugs on hematopoietic cells, veterinary practice, and even the study of new strategies in cellular therapy for some human diseases.     

Corneal endothelium of the Magellanic penguin (Spheniscus magellanicus) by scanning electron microscopy.

The corneal endothelium is essential for the maintenance of the corneal transparency. The aim of this study was to examine the morphology of the endothelial surface and perform morphometric analysis of the normal corneal endothelial cells of the Magellanic penguin (Spheniscus magellanicus) using scanning electron microscopy. The present work demonstrates that the corneal endothelium of the Magellanic penguin is similar to those described in other vertebrates.