Density‐dependent prophylaxis in primary anti‐parasite barriers in the velvetbean caterpillar

1. Organisms rely on a set of primary barriers to prevent invasion by parasites, and secondary defences to fight parasites that breach the primary barriers. However, maintaining these defences to be active and effective is costly. Thus, hosts increase investment in anti‐parasite defences under situations of high risk of infection and reduce defences when the risk is reduced (the ‘Density‐Dependent Prophylaxis’ hypothesis). 2. In the present study, it was tested whether the midgut primary defences of the velvetbean caterpillar Anticarsia gemmatalis Hübner present density‐dependent plasticity, and also whether these defences could be induced by a viral pathogenic challenge. The aim was to examine whether morphometry and the structure of the midgut and peritrophic matrix (PM) change in accordance with colour transition in caterpillars, and whether such changes may provide the caterpillars a more protective barrier against invasion by Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV). 3. It was found that PM and the midgut epithelium of the velvetbean caterpillar change plastically according to phenotype, itself a response to changes in population density. Caterpillars reared at high densities (black phenotype) had a considerably thicker midgut epithelia and peritrophic matrices than those reared individually (green phenotype), and there was also more chitin in the PM of the former. 4. This was interpreted as the first demonstration of increased investment in primary, barrier, defences against parasites, in response to increased conspecific density and an increased risk of infection. The possibility that this arises as a positive result of pleiotropy is discussed further, wherein the biochemical pathways responsible for the up‐regulation of the immune system are also involved in midgut properties.     

The effect of adenine nucleotides and inhibitors of ADP translocation on uncoupled electron flux in bean mitochondria

The uncoupled electron flux and the influence of adenine nucleotides on this flux in mitochondria isolated from hypocotyls of Vigna sinensis (L.) Savi cv. Seridó were examined. In order to avoid the functioning of other enzymes capable of utilizing adenine nucleotides the reaction medium was free of Mg²⁺. When an oxidizable NADH -linked substrate such as L-malate was used, a stimulatory of adenosine-5′-monophosphate (AMP) and adenosine-5′-diphosphate (ADP) on uncoupled respiration was manifested. The stimulatory effect of AMP and ADP could not be shown when succinate was the substrate. Atractyloside and carboxyatractyloside had no effect on the stimulatory role played respectively by AMP and ADP in the presence of carbonyl cyanide p-trifluoromethoxy-phenylhydrazone (FCCP).     

Herpetomonas roitmani (Fiorini et al., 1989) N. Comb.: A Trypanosomatid with a Bacterium-like Endosymbiont in the Cytoplasm

The trypanosomatid previously described as Crithidia roitmani is characterized here at the ultrastructural and biochemical levels. The data indicates that the parasite belongs to the Herpetomonas genus, and we therefore suggest the flagellate to be denominated as Herpetomonas roitmani n. comb. Cladistic analysis of isoenzyme data generated by eight different enzymes showed that the parasite presented a distinct banding pattern and could be grouped with some Herpetomonas spp., but not with Crithidia spp., used as reference strains. Accordingly, when the parasites were grown for longer periods in Roitman's defined medium, expontaneous differentiation from promastigotes to opisthomastigotes (typical of the Herpetomonas genus) occurred. Transmission electron microscopy revealed the presence of bacterium-like endosymbionts in the cytoplasm of all evolutive forms of the parasite. All morphological alterations characteristic of endosymbiont-bearing trypanosomatids could be observed.     

Short-Term In Vitro Culture and Molecular Analysis of the Microsporidian, Enterocytozoon bieneusi

ABSTRACT. The microsporidium, Enterocytozoon bieneusi , causes a severe, debilitating, chronic diarrhea in patients with the acquired immunodeficiency syndrome. Specific diagnosis of intestinal microsporidiosis, especially due to Enterocytozoon , is difficult and there is no known therapy that can completely eradicate this parasite. Preliminary studies indicate that a short term (about 6 months) in vitro culture of this parasite yielding low numbers of spores, may be established by inoculating human lung fibroblasts and/or monkey kidney cell cultures with duodenal aspirates and or biopsy from infected patients. The cultures may subsequently be used for the isolation and molecular analysis of parasite DNA.     

Gram-Chromotrope: a New Technique that Enhances Detection of Microsporidial Spores in Clinical Samples

We have developed a new staining procedure that combines the traditional Gram staining for bacteria and the Weber's chromotrope staining method, the standard technique for the detection of microsporidia spores in clinical Specimens. This “Gram-chromotrope” staining technique enhances the staining characteristics of microsporidia spores and facilitates the easy detection and differentiation of spores from other microorganisms that are found in clinical specimens, especially stool samples. This new technique is fast, reliable, and simple to perform, and can be easily adapted for use in clinical laboratories.     

Phylogenetic relationships among marsupial frog genera (Anura: Hylidae: Hemiphractinae) based on evidence from morphology and natural history

We reassess the phylogenetic relationships of genera of hemiphractine hylid frogs (Marsupial Treefrogs) and discuss the evolution of several distinctive characters within this group using parsimony analysis. Fifty-one morphological and life-history characters were sampled from two species of Cryptobatrachus , three species of Flectonotus , 17 species of Gastrotheca , all five species of Hemiphractus , and one species of Stefania as the ingroup and three hyline, one phyllomedusine, and one pelodryadine species as outgroups. Our results support the mon-ophyly of Flectonotus, Cryptobatrachus , and Hemiphractus. Gastrotheca is paraphyletic with respect to Hemiphractus , dorsal pouches were lost in the ancestor of Hemiphractus. Direct development is a synapomorphy for Hemiphractinae and tadpoles were regained independently several times. These results stand in stark contrast to the prevailing paradigm regarding marsupial frog relationships.     

Changes in Cell Surface Anionogenic Groups Induced by Propranolol in Herpetomonas muscarum muscarum

The effects of propranolol (10⁻³ mM) on the surface anionic groups of Herpetomonas muscarum muscarum were analysed by cell electrophoresis, by ultrastructural cytochemistry and by identification of sialic acids using paper chromatography. Differentiation of H. muscarum muscarum induced by propranolol treatment caused a significant increase in the net negative surface charge. Binding of cationized ferritin (CF) and colloidal iron hydroxide particles was observed at the cell surface of both untreated and propranolol-treated cells. In cells incubated in the presence of the drug the CF particles were distributed in all membrane regions. However, there were small areas where the particles were absent. In H. muscarum muscarum exposed to propranolol the density of residues of sialic acid per cell was higher, and the agglutinating activity with Sendai virus was more intense. However, the pattern of sialic acid, characterized by the presence of N-acetylneuraminic acid derivative, was not modified upon cell interaction with the drug. Treatment of both control and propranolol-treated protozoa with neuraminidase significantly reduced the surface charge. These findings suggest that sialic acid residues are the major anionogenic groups exposed on the surface of H. muscarum muscarum .