Resistance to the cereal cyst nematode (Heterodera avenae Woll.) transferred from the wild grassAegilops ventricosa to hexaploid wheat by a “stepping-stone” procedure

Transfer of resistance toHeterodera avenae, the cereal cyst nematode (CCN), by a stepping-stone procedure from the wild grassAegilops ventricosa to hexaploid wheat has been demonstrated. The number of nematodes per plant was lower, and reached a plateau much earlier, in the resistant introgression line H93-8 (1–2 nematodes per plant) than in the recipient H10-15 wheat (14–16 nematodes per plant). Necrosis (hypersensitive reaction) near the nematode, little cell fusion, and few, often degraded syncytia were observed in infested H93-8 roots, while abundant, well-formed syncytia were present in the susceptible H10-15 wheat. Line H93-8 was highly resistant to the two Spanish populations tested, as well as the four French races (Fr1-Fr4), and the British pathotype Hall, but was susceptible to the Swedish pathotypes HgI and HgIII. Resistance was inherited as though determined by a single quasi-dominant factor in the F₂ generations resulting from crosses of H93-8 with H10-15 and with Loros, a resistant wheat carrying the geneCre1 (syn.Ccn1). The resistance gene in H93-8 (Cre2 orCcn2) is not allelic with respect to that in Loros. RFLPs and other markers, together with the cytogenetical evidence, indicate that theCre2 gene has been integrated into a wheat chromosome without affecting its meiotic pairing ability. Introduction ofCre2 by backcrossing into a commercial wheat backgroud increases grain yield when under challenge by the nematode and is not detrimental in the absence of infestation.     

Kinetic study of the cytotoxic effect of α-sarcin,a ribosome inactivating protein fromAspergillus giganteus,on tumour cell lines: protein biosynthesis inhibition and cell binding

-Sarcin is a ribosome inactivating protein produced by the mouldAspergillus giganteus. The effect of this protein on eight different tumour cell lines has been studied in the absence of any agent affecting membrane permeability. The protein is cytotoxic for all the tumour cell lines considered. -Sarcin modifies the cell proliferation pattern by inhibiting the protein biosynthesis of the cultured cells. No membrane damage produced by -sarcin has been observed by measuring lactic dehydrogenase leakage. Alteration on the cell mitochondrial activity has not been detected upon treatment with -sarcin. Differences on the extent of the protein binding to the cells have been observed by flow cytometric measurements. The kinetic analysis of the protein biosynthesis inhibition produced by -sarcin reveals an -sarcin concentration-dependent lag phase followed by a first order decrease of the protein synthesis rate. This parameter is dependent on the external -sarcin concentration. A saturable component for the action of -sarcin is also deduced from these experiments. Results are discussed in terms of the protein passage across the cell membrane as the potential rate-limiting step for the action of -sarcin.     

In vitro susceptibility ofPityrosporum ovale (Malassezia furfur) to human androgenic steroids

Cells ofPityrosporum ovale that colonize human pilosebaceous units are constantly exposed to cutaneous androgenic steroids. The aim of our study was to find out whetherP. ovale is susceptible to these hormones. Three strains ofP. ovale were grown in vitro in the presence of various concentrations oftestosterone, dehydroepiandrosterone, androstenedione, androstanedione, 5--dihydrotestosterone andprogesterone (10, 100, and 1000 µg/ml; agar dilution assays). In addition, three strains ofCandida albicans were also exposed to equal concentrations of the same androgens. As a result, allP. ovale strains were suppressed by 1000 µg/mlandrostenedione, which was the strongest inhibitor. The other androgenic steroids also significantly reducedP. ovale growth at different concentrations, depending on the hormone used and the strain tested.Progesterone was inhibitory at the highest concentration for oneP. ovale strain only.Candida albicans was not affected by any of the androgens. These findings demonstrate an in vitro susceptibility ofP. ovale to high concentrations of human androgenic steroids. A relevance of this interaction for the in vivo fungus-host relation is not apparent.     

Pythium aphanidermatum root rot of pawpaw (Carica papaya L.) in Nigeria

Root rot of pawpaw (Carica papaya L.) reported in Nigeria is caused byPythium aphanidermatum which was consistently isolated from diseased plant parts and highly pothogenic. Out of 16 different media tested, it grew best on corn-meal-agar (CMA) and CMA supplemented with cellulose and sucrose. The highest number of oospores/ml was on CMA with average diameter of 19.9±0.1 µm. The symptom is characterized by dark brown rot of roots, absence of secondary roots and disintegration of internal tissue of the main root. These cause the progressive decline of the aerial parts of the tree untill it dies.     

Evidence for growth ofSporothrix schenckii on dead but not on living sphagnum moss

When clinical isolates ofSporothrix schenckii were inoculated onto the apices of living or dead sphagnum moss plants maintained under growth chamber conditions, populations of the fungus, assessed by standard dilution plate methods, increased swiftly up to about 70-fold on moist, dead plants but did not increase on the live moss. Light and scanning electron microscopy revealed fungal growth and sporulation on and within dead plants, but no evidence of either on live plants. These data provide indirect support for the contention thatS. schenckii does not grow on living sphagnum in bogs, but rather that sporotrichosis epidemics associated with sphagnum moss are likely to result from contamination of the dead plants at some point(s) in the chain of events during or after harvest. One practical implication of our results is that precautions should be taken to insure that sphagnum moss is stored dry and that it is not wetted any sooner than necessary before use. We also report here improvement of the Mycose isolation medium by an increase in cycloheximide from 400 to 800 mg/l, chloramphenicol from 50 to 250 mg/l, and the addition of rifampicin at 20 mg/l.     

Studies on the fungi associated with tomato fruit rots and effects of environment on storage

Seven fungi associated with fruit rot of tomato were isolated includingFusarium equiseti, F. chlamydosporum, Alternaria solani, Geotrichum candidum, Acremonium recifei, Aspergillus flavus andA. niger. They were all pathogenic on tomato fruits, most pathogenic beingGeotrichum candidum followed byA. niger. Least rot was caused byAlternaria solani. The optimum temperature for maximum rotting caused byG. candidum, A. niger andA. flavus was 30°C. The relative humidity for maximum rot ranged from 70–90%. Tomato fruits stored well at 0–10°C and rather poorly at 20–30°C. Fruits stored at 35°C showed blemishes. The best RH for storage ranged between 60 and 90%.     

Histoplasma capsulatum peritonitis associated with continuous ambulatory peritoneal dialysis

This paper reports a case of infection byHistoplasma capsulatum apparently restricted to the peritoneum in a woman submitted to continuous ambulatory peritoneal dialysis. Diagnosis was established by culture of dialysis fluid and peritoneal nodule and by histopathologic examination.     

Effect of glucose starvation on germ-tube production byCandida albicans

By incubating starved and unstarved yeast cells in synthetic media with a pH of 4.5 or 6.7 at 37°C the effect of a 3 hours' glucose starvation on germ-tube production byCandida albicans was evaluated. In addition the endocellular content of total carbohydrates, glycogen, trehalose and proteins after and before the starvation were dosed. The most interesting result was the overcoming of the pH-regulated dimorphism, thanks to the starvation treatment. Infact the starved cultures produced germtubes indifferently in neutral or acid media, whereas the filamentation of the unstarved cultures was more copious in pH 6.7 medium. The endocellular content of trehalose and protein was unchanged, whereas total carbohydrates and glycogen showed a shortage after the 3 hours' glucose starvation. The possible involvements of these metabolic changes in the regulation of dimorphic transition are discussed.     

A potent specific inhibitor of 6-phosphogluconate dehydrogenase ofCryptococcus neoformans and of certain other fungal enzymes

A particular lot of the zwitterionic buffer, 2(N-morpholino) ethane sulfonic acid (MES), contained a contaminant that inhibited a number of fungal NADP-dependent dehydrogenases. Enzymes that were particularly sensitive include 6-phosphogluconate dehydrogenases fromCryptococcus neoformans andSchizophyllum commune and glucose-6-phosphate dehydrogenase fromSchizophyllum commune. A number of NADP-dependent dehydrogenases of animal origin were tested and all were completely insensitive to inhibition except for rat liver 6-phosphogluconate dehydrogenase, which was 10-fold less sensitive than theCryptococcal enzyme. The pattern of inhibition in all cases was linear competitive versus NADP. The inhibitor has been purified and identified as an ethylenesulfonic acid oligomer. This inhibitor holds promise as a model compound for the development of a specific antifungal agent.     

Dialysable leukocyte extracts modify the course of experimental paracoccidioidomycosis in the Syrian hamster

The effect of dialysable leukocyte extracts (DLE) obtained from hamsters immunized withParacoccidioides brasiliensis (immune DLE) and from non-immunized hamsters (non-immune DLE) was studied in hamsters inoculated withP. brasiliensis by the intratesticular route. Treatment with immune or non-immune DLE was started during the third week of infection and was repeated at 7, 11, 15 and 19 weeks. A group of untreated infected animals was used as control. Animals were submitted to the delayed hypersensitivity skin test toP. brasiliensis antigen (PbAg) in vivo and assayed in vitro by the macrophage migration inhibition test in the presence of Phytohemagglutinin (PHA) and PbAg and by immunodiffusion for specific antibody. The animals were sacrificed at 4, 8, 12, 16 and 20 weeks. The morphology and extension of the lesions were studied at the inoculation site, and in lymph nodes, lungs, liver, spleen and kidneys. In contrast to the controls, animals treated with both DLEs maintained a positive cell-mediated immune response throughout the experiment and developed less extensive infection with a significantly lower number of fungi in the lesions. The results suggest that immune and non-immune DLE preparations modified the evolution of experimental paracoccidioidomycosis with equal efficiency. This similarity may be explained by the immunoregulatory activities of both extracts.