Fever and acute phase response induced in dwarf goats by endotoxin and bovine and human recombinant tumour necrosis factor alpha

Tumour necrosis factor (TNF), a polypeptide produced by mononuclear phagocytes, has been implicated as an important mediator of inflammatory processes and of clinical manifestations in acute infectious diseases. To study further the potential role of TNF in infectious diseases, recombinant Escherichia coli (E. coli) derived human (r.HuTNF-alpha) and bovine TNF (r.BoTNF-alpha) were intravenously (i.v.) administered in dwarf goats. Rectal temperature, heart rate, rumen motility, plasma zinc and iron concentrations, and certain other blood biochemical and haematological values were studied and compared with the changes seen after E. coli endotoxin (LPS) was administered (dose: 0.1 microgram/kg i.v.). Following a single injection of 4 micrograms/kg of r.BoTNF-alpha, shivering and biphasic febrile response were observed, accompanied by tachycardia, inhibition of rumen contractions, drop in plasma zinc and iron concentrations, lymphopenia, and neutropenia followed by neutrophilia. The i.v. administration of a single injection of 4 micrograms/kg r.HuTNF-alpha induced shivering and biphasic febrile responses, accompanied by anorexia and a similar drop in plasma trace metal concentrations when compared with r.BoTNF-alpha-treated goats. The TNF-alpha-induced symptoms were essentially the same as those that occurred after LPS administration. However, the time of onset of these changes after the injection of TNF-alpha was significantly shorter than after LPS. Moreover, the r.BoTNF-alpha induced a longer lasting neutrophilic leucopenia, less neutrophilia, and a more persistent lymphopenia than after LPS injection. Neither r.BoTNF-alpha nor LPS caused severe haemo-concentration. Furthermore, no cross-tolerance between r.BoTNF-alpha and LPS could be demonstrated. We conclude that both r.BoTNF-alpha and r.HuTNF-alpha induce many of the physiologic, haematologic and metabolic changes that characterize the acute phase response to LPS. The overlapping biological activities of r.BoTNF-alpha, r.HuTNF-alpha and LPS in dwarf goats may indicate that both recombinant tumour necrosis factors have some homology with caprine TNF-alpha.     

Effects of triazole fungicides on sterol biosynthesis during spore germination of Botrytis cinerea,Venturia inaequalis and Puccinia graminis f. sp. tritici

With three plant pathogens,Botrytis cinerea, Venturia inaequalis and Puccinia graminis f. sp.tritici, the time course of sterol biosynthesis during spore germination was examined by labeling experiments along with the question whether this pathway could be inhibited by triazole fungicides. Conidia ofB. cinerea andV. inaequalis are able to synthesize sterols immediately after the beginning of the germination process when the germ tubes have not yet emerged. On the contrary uredospores ofP. graminis start sterol biosynthesis after 6 to 8 h germination time almost at the end of the germ tube phase, indicating that sterol reserves of the spores are likely to be used for the germ tube growth.The sterol C-14 demethylation appeared to be the rate limiting step within the sterol biosynthetic pathway: the half life of 24-methylenedihydrolanosterol was less than 1 h forB. cinerea. It was more than 1 h forV. inaequalis and 3 h forP. graminis. Independent of these differences in the time course of sterol biosynthesis and in the C-14 demethylation rate, the synthesis of sterols in germinating spores was strongly inhibited by triazole fungicides in all three pathogens examined. In contrast toP. graminis, this inhibition could be demonstrated withB. cinerea andV. inaequalis even in ungerminated conidia, indicating that the fungicides were rapidly taken up and reached their target within 1 or 2 h. These results are discussed along with the question whether spore germination can be used as a bioassay for the estimation of sensitivities of triazole fungicides.     

In vitro selection of sterol-biosynthesis-inhibitor (SBI)-resistant mutants in Monilinia fructicola (Wint.) Honey

The inherent resistance risk forMonilinia fructicola against sterol-biosynthesis inhibitors (SBIs) was estimated inin vitro andin vivo laboratory studies. Several mutant strains were selected on media amended with the triazole fungicides penconazole, etaconazole or the morpholine fungicide fenpropimorph.The potential forM. fructicola to develop resistance to the triazoles or to the morpholines was similar.The level of resistance attained did not differ for the two classes of fungicides after a single cycle of treatment with nitrosoguanidine (NTG). Attemps to select mutants with a higher level of resistance to penconazole after successive mutagenic treatments were successful. Most of the mutants were less fit than wild-type strains. Mutants with a low level of resistance had an almost normal mycelial growth rate, whereas growth of mutants with a higher level of resistance was significantly reduced. Spore production was highest in the wild-type strains, similar to the latter in a few resistant strains and less in most others. Only one mutant with an intermediate level of resistance could successfully compete in a mixed population with a wild type strain during successive infection cycles on peaches. Resistance was not stable in highly resistant mutants. Cross resistance to the inhibitors of 14-methylsterol demethylation (DMIs) tested was confirmedin vitro andin vivo for all mutant strains. One DMI-resistant mutant was also resistant to fenpropimorph and two fenpropimorph-resistant mutants were resistant to penconazole.     

Colonization of rose by Sphaerotheca pannosa (Wallr.) Lév. var. rosae Wor.

Colonization of rose by powdery mildew (Sphaerotheca pannosa) is described in terms of mycelium growth, conidiophore production and sporulation in time. The data used are gathered during different years, put together and treated by means of graphic models. Colonies could be separated into fast and slow growing colonies. Colonies initiated on leaves of increasing age showed a decreasing growth rate. Production of conidiophores and conidia started on the same day, and the relative activity of conidiophore production reached its maximum 6 days after the end of the latency period, followed 1 day later by the maximum activity of conidium production. Both conidiophore and conidium production continued for a long time at a low level. The effect of leaf age on conidiophore production found expression in differences in production rate during the first days of colony development and in final production levels. Observations on naturally infected leaves in an outdoor experiment showed a rapid decrease of sporulation on leaves of 10 days and older. Highest percentages of sporulating leaf area were observed on leaves between 7 and 10 days old.     

Inactivation of soil-borne plant pathogens during small-scale composting of crop residues

Samples of heavily infested crop residues were incorporated in static compost heaps (2.5–4.6 m³) of the Indore type. Temperature increased to 50–70°C within 6 days depending on the type of crop residues used and the location within the heap. The heat phase (>40 °C) lasted 2–3 weeks and was followed by a c. 5-months maturation phase (<40 °C). Among the 17 pathogens tested, onlyOlpidium brassicae and one of the four formae speciales ofFusarium oxysporum that were tested survived composting, but also their inoculum was greatly reduced.Survival during specific phases of composting was studied by incorporation and retrieval of samples at various stages of the process.F. oxysporum f. sp.melonis was completely inactivated andO. brassicae andPlasmodiophora brassicae were almost completely inactivated during the short heat phase. The three pathogens survived the long-lasting maturation phase without loss of viability. Heat evolved during composting was found to be the most important factor involved with sanitation of crop residues. The possible involvement of fungitoxic conversion products and microbial antagonism is discussed.