Production of microsporidia pathogenic to the Colorado potato beetle (Leptinotarsa decemlineata) in alternate hosts

The microsporida Nosema gastroideae and N. equestris, which are highly pathogenic for Leptinotarsa, have been successfully produced in some other chrysomelid species, Gastrophysa polygoni and G. viridula. As the principal target host, Leptinotarsa is very susceptible to these pathogens, and death occurs before massive sporulation by the microsporidia. By contrast, the infected larvae of G. polygoni or G. viridula are able to develop until the adult stage when most of the tissues become filled with spores. In addition, the larvae and adults of these species can be reared in the laboratory on Polygonum aviculare and Rumex obtusifolius. These plants have longer vegetative periods and are better sources of food than potato leaves. In both species of Gastrophysa the yields of spores related to unit weight were about five times higher than in Leptinotarsa. In the adults of G. viridula there was up to 4.8 × 10⁶ spores mg^?1 body weight of N. gastroideae, or 9.1 × 10⁶ spores mg^?1 of N. equestris. The higher content of microsporidian spores facilitates their purification and isolation.     

Respiration and microflora of the rhizosphere soil of wheat influenced by foliar application of urea

The numbers of micromycetes and bacteria were investigated with respect to oxygen consumption in the rhizosphere soil of wheat and in non-rhizosphere soil. Plants after foliar application of urea (2 % solution) and non-treated plants were cultivated in degraded chernozem and garden soil in a green-house. Changes in oxygen consumption by the suspensions of rhizosphere and non-rhizosphere soils corresponded to changes in the number of bacteria designated as the rhizosphere effect (R/S). Values of R/S depended on the presence of organic substrates. Changes in oxygen consumption by the soil suspension from the rhizosphere of wheat occurring due to foliar application of urea corresponded to changes in the amount of microflora. The results obtained are discussed with respect to a possible utilization of the data to follow metabolic activity of soils in a natural environment (in situ) determined according to oxygen consumption by a soil suspension, and to assess changes in the microflora of rhizosphere and non-rhizosphere soil.     

Passage of foreign proteins across the intestinal wall of newborn rabbits

Foreign proteins penetrate across the wall of rabbit jejunum takes place within the first eight hours after birth. Sensitization by foreign protein (cow's milk protein) during early postnatal ontogeny is the cause of death of artificially fed germfree rabbits at the age of 21-23 d.     

Translation of poly(U) on ribosomes from Streptomyces aureofaciens

Slowly cooled cells of Streptomyces aureofaciens contained mainly tight-couple ribosomes. Maximum rate of polyphenylalanine synthesis on ribosomes of S. aureofaciens was observed at 40°C, while cultures grew optimally at 28°C. Ribosomes of S. aureofaciens differed from those of E. coli in the amount of poly(U) required for maximum synthetic activity. The polyphenylalanine-synthesizing activity of E. coli ribosomes was about 3-times higher than that of S. aureofaciens ribosomes. The addition of protein S1 of E. coli or the homologous protein from S. aureofaciens had no stimulatory effect on the translation of poly(U). In order to localize alteration(s) of S. aureofaciens ribosomes in the elongation step of polypeptide synthesis we developed an in vitro system derived from purified elongation factors and ribosomal subunits. The enzymatic binding of Phe-tRNA to ribosomes of S. aureofaciens was significantly lower than the binding to ribosomes of E. coli. This alteration was mainly connected with the function of S. aureofaciens 50 S subunits. These subunits were not deficient in their ability to associate with 30 S subunits or with protein SL5 which is homologous to L7/L12 of E. coli.     

The problem of carotenoid biosynthesis in the taxonomy of genera Rhodotorula and Rhodosporidium

This report presents results in the composition of major carotenoids of various coloured mutants of the genusRhodotorula and of mating types of the genusRhodosporidium. The separation of carotenoid intermediates was carried out by thin layer chromatography using Silufol 254 Kavalier and the determination of eluated spots by spectrophotometry. There was found no difference in carotenoid composition of both mating typesa and α of individual species of the genusRhodosporidium. The vegetative and sexual reproduction ofRhodotorula andRhodosporidium can be separated from the carotenogenesis using 10^?4 mol diphenylamine. It was concluded that lycopene could be the intermediate to mono- and dicyclic carotenoids; in the case of partial inhibition of the dehydrogenation step the direct cyclization of neurosporene to β-zeacarotene can be expected. An unknown compound, probably lycopersene was found and was considered to be the precursor of phytoene. Phytoene and phytofluene were proved in all studied samples. Nutritional conditions (vitamins, sulfur amino acids, etc.) are able to shift the ratios between major carotenoids. Rhodotorula aurantiaca strains were observed to be auxotrophic mutants of various characters and the existence of this species as independent one, was denied.     

Histochemical demonstration of the intestinal hetero-β-galactosidase (glucosidase)

Summary The histochemical demonstration of hetero--galactosidase (glucosidase) has been attempted in sections and zymograms of rabbit, monkey and human intestine and of rat kidney.The leakage of this enzyme from unfixed sections was prevented by the use of cold microtome sections adherent to semipermeable membranes. Methods with -D-glucosides and galactosides of 6-Br-2-naphthol (postincubation azocoupling with Fast Blue B as well as simultaneous azocoupling with hexazonium-p-rosaniline), of -naphthol (simultaneous azocoupling with hexazonium-p-rosaniline) and of 4-Cl-5-Br-3-indolyl (with ferricyanide, phenazonium methosulfate or nitro BT and without any oxidation agent) were used an evaluated concerning the specificity, localization ability and inhibition of enzyme activity. Pretreatment of sections with distilled water or saline and inhibition by p-Cl-mercuribenzoate, glucono- and galactono-lactones were used for the characterization of the demonstrated enzyme activity.6-Br-2-naphthyl--D-glucoside is the most specific substrate for hetero--galactosidase. It is not split by lactase and acid -galactosidase. Only lysosomal -glucosidase can interfere. Because the latter enzyme is membrane-bound the difference in color intensity between untreated and prewashed sections are due to hetero--galactosidase. Only localization on the cellular (not intracellular) level can be achieved, however.The simultaneous azocoupling method with -naphthyl--D-glucoside and hexazonium-p-rosaniline enables a very good localization of hetero--galactosidase in the rabbit intestine. Due to a great inhibition exerted by hexazonium-p-rosaniline on the enzyme activity the method is unsuitable for the detection of hetero--galactosidase in zymograms and in the human intestine. Interference of lactase (or lactase-phlorizine hydrolase complex) is to be considered. The lysosomal -glucosidase does not seem to interfere.Indigogenic methods are not sensitive either. With ferricyanide as an oxidation agent it was not possible to detect the activity of hetero--galactosidase in zymograms and in sections. This is possibly due to overoxidation of indigo. The same holds true for phenazonium methosulfate used for the processing of zymograms. However, it was possible to reveal the activity of hetero--galactosidase in sections of the rabbit and monkey intestine with phenazonium methosulfate as oxidation agent. Nitro BT enhanced the coloration both in zymograms and in sections. In the latter case diffusion artifacts cannot be prevented, however. The interference of lactase, lysosomal -galactosidase and possibly of lysosomal -glucosidase (depending on the glycoside used) is always to be considered.Hetero--galactosidase was localized in the cytoplasm (particularly in the supranuclear region) of differentiated enterocytes covering the villi of the rabbit (the highest activity), monkey and human (the lowest activity) intestine. In crypt enterocytes and in cells of Brunner's glands the activity was lower. The occurrence of a low activity of hetero--galactosidase in the brush border of enterocytes of the rabbit intestine was also demonstrated.A proximodistal gradient was observed in the rabbit and monkey intestine, the upper jejunum displaying the highest activity.In jejunal biopsies of patients with celiac sprue (in the acute stage of the disease) the activity of hetero--galactosidase was lowered. No changes of activity were observed in jejunal biopsies of patients with isolated deficiencies of lactase or sucrase.In the rat kidney the enzyme was demonstrated particularly in the cytoplasm of cells of proximal convoluted tubules.     

Antibacterially active substances

Synthesis of two hydroxy-derivatives of nalidixic acid as a result of microbial transformation was demonstrated in certain species of the genusAspergillus. Aspergillus alliaceus produced 7-hydroxy-nalidixic acid andAspergillus niger 6-hydroxy-nalidixic acid. It was demonstrated that the antibacterial activity of both hydroxy-derivatives (tested inEscherichia coli) was lower than that of the initial nalidixis acid.