Biological properties of opportunistic enterobacteria isolated from healthy persons and from patients with intestinal dysbacteriosis

The comparative study of the signs of pathogenicity in 724 enterobacterial strains and in some species of nonfermenting microorganisms isolated from the feces of 115 somatic patients with intestinal dysbacteriosis and 80 healthy persons (565 strains) has revealed that microorganisms belonging to the same species may essentially differ in their occurrence, in the activity of the enzymes (DNAase, RNAase, phosphatase) which they release into the culture medium, as well as in their capacity to give the positive reaction with Congo red. At the same time, the cultures isolated in cases of intestinal dysbacteriosis have been found to be biologically more active.     

Biological properties of opportunistic enterobacteria isolated from the blood of patients

The comparative study of the signs of pathogenicity in enterobacteria (119 strains) isolated from the blood of 145 patients with the clinical symptoms of sepsis and from the feces of healthy persons (560 strains from 220 persons) has demonstrated that the same species of opportunistic microorganisms may essentially differ in the formation of DNase, RNase, as well as in their capacity for the positive reaction with Congo red. The possibility of using the above-mentioned signs of pathogenicity for diagnostic purposes as additional signs for the differentiation of virulent cultures of opportunistic enterobacteria from avirulent ones is suggested.     

The biological properties of Bifidobacterium]

The possibility of the formation of exoenzymes, such as DNAase, RNAase and hemolysin, by bifidobacteria was studied in comparison with their acid-forming and adhesive activity. Bifidobacterium reference strains, originally isolated from healthy adults and children, were studied. The study involved altogether 73 strains of bifidobacteria, including 24 B. bifidum strains, 13 B. adolescentis strains, 7 B. infantis strains, 10 B. breve strains and 19 B. longum strains. The bifidobacteria under study were shown to differ not only in the presence and activity of properties useful for macroorganisms, but also in the presence of enzymes having depolymerizing activity (DNAase, hemolysin). Thus, out of 73 strains under study 9 proved to be DNAase-positive and 6, hemolysin positive. At the same time a specific feature of bifidobacteria was their high acid-forming activity with the complete absence of RNAase activity and insignificant DNAase- and hemolysin-forming activity.     

The microflora of ulzerous zone mucosa in patients with duodenal ulcer

Bacteriological study of the biopsies taken from gastric and duodenal mucosa of 10 healthy volunteers and 74 patients with duodenal ulcer, was carried out. In the gastroduodenal zone of healthy subjects microorganisms of 6 genera (Streptococcus, Candida, Staphylococcus, Bacillus, Helicobacter and Lactobacillus) were detected. H. pylori was isolated in 20% of cases only in biopsy specimens taken from the antral section of the stomach of healthy as monoculture or in combination with C. albicans. In patients with duodenal ulcer activation of opportunistic microflora was observed in the periulcerous zone. More often H. pylori occurred in associations with fungi of the genus Candida, streptococci, staphylococci, enterobacteria, Pseudomonas and other microorganisms (of more than 30 genera). Quantitatively the dominating microorganisms (3.8-5.7 lg CFU/g) were H. pylori, fungi of the genus Candida, bacteria of the genera Streptococcus, Peptostreptococcus, Bacteroides, Gemella, Prevotella, Veillonella, Peptococcus, Bacillus, different species of opportunistic enterobacteria, as well as bacteria of the genera Staphylococcus, Micrococcus, Corynebacterium, Neisseria, Pseudomonas, etc. Opportunistic bacteria detected in the ulcerous zone, as a rule, expressed hemolytic, lecithinase, RNAase, caseinolytic, catalase and urease activity. Sonicated filtrates of such cultures produced a cytotoxic effect on cells HEp-2. Ulcer is an infected wound that needs sanitation.     

Study of the role of conditionally pathogenic enterobacteria in acute intestinal diseases]

Conditionally pathogenic enterobacteria were isolated in 34.2% of the patients with acute intestinal diseases (1559 cases examined in all). Such representatives of enterobacteria as Klebsiella, Hafnia, Citrobacter of 4, 8, and 22 serological groups. Proteus mirabilis of 6, 10, 13, 26, and 28 serological groups, and Proteus morganii mostly played the etiological role. In some of the patients conditionally pathogenic microbes only aggravated the main disease, taking part in the development of dysbacteriosis. In healthy individuals these microbes were much more rare and were encountered inconstantly.     

Various biological properties of enterobacteria, isolated from patients with diarrhea

381 enterobacterial strains isolated from patients with acute enteric diseases were studied. Of these, 279 strains, as well as 20 strains isolated from 50 healthy children, were studied for the presence of adhesins, hemolysins and catalase. The comparison of the hemagglutinating activity of enterobacteria isolated from sick and healthy children revealed no essential differences between them. 15.8% of enterobacterial strains isolated from sick children possessed hemolytic activity, while strains isolated from healthy children did not induce the hemolysis of erythrocytes. All enterobacterial strains isolated from patients with acute enteric diseases were multi-resistant to antibiotics. A multitude of different antibiograms was obtained, most of them occurring only once. In 1985 the number of multiresistant (i. e. resistant to 11 and more antibiotics) strains dropped from 61% to 26..9% in comparison with 1981.     

Interaction of a causative agent with associative bacteria during salmonellosis

AIM: To determine a composition of gut microflora during salmonellosis and to study the modification of persistent characteristics (antilysozyme activity, ALA) of symbiotic microorganisms in associations. MATERIALS AND METHODS: Bacteriologic study of feces was performed in 90 patients aged 18-39 years, which were divided to three groups: patients with salmonellosis in acute phase, reconvalescent patients, and conditionally healthy persons. Condition of gut microflorawas determined; microorganisms associated with Salmonella infection were isolated, and their influence on ALA of Salmonella was studied. RESULTS: Gut microbiocenosis was more diverse in patients compared with healthy persons. Significant reduction of bifidobacteria quantity (to 10(7) CFU/g of feces and less), especially in reconvalescent period, was noted. Association between bifidoflora deficiency and excessive increase of quantity of yeast fungi was revealed. It was determined that exometabolites of indigenous anaerobic microflora (bifidobacteria) promoted decrease of ALA of Salmonella, whereas opportunistic facultatively anaerobic microorganisms (enterobacteria, staphylococci) rendered mainly stimulating effect on the ALA of Salmonella. CONCLUSION: Obtained data reveal characteristics of bacterial interactions in associative symbiosis and provide insights about mechanisms of formation of pathobiocenosis and state of bacterial carriage.     

Fecal microflora in healthy young people

The study of the intestinal microflora in 119 young adults was carried out. A high content of anaerobic representatives of the intestinal microflora (bifido- and lactobacteria) and extremely wide fluctuations in the number of E. coli (1-5 million to 700-800 million cells per g of feces) were shown. The species composition of the facultative group was found to be variegated. Staphylococci, yeast, fungi, opportunistic enterobacteria, as well as Escherichia and cocci with changed characteristics were detected. 23.5% of the subjects showed a high content of E. coli (greater than 200 million cells per g of feces) accompanied by the increased occurrence of Klebsiella and Escherichia with changed properties. These persons can be regarded as a high risk group with a higher incidence of acute intestinal diseases with unknown etiology.     

Detection of enterotoxigenic enterobacteria with adhesion antigens in acute intestinal diseases

As revealed in this study, 56.8 +/- 4.3% of children under 1 year, suffering from acute intestinal diseases of unknown (i.e. not determined by common bacteriological methods) etiology, show the presence of enterobacteria capable of the combined synthesis of enterotoxins and adhesion antigens. No such cultures are isolated from healthy children. In cases of diarrhea of domestic animals (piglets and calves), frequent isolation of enterobacteria characterized by both, toxigenicity and capacity for producing adhesion antigens (50.6 +/- 4.8% and 42.9 +/- 4.8% respectively), is noted.     

Characterization of alkaline nuclease from rat liver mitochondria.

The alkaline nuclease (pH optimum 9.0) has been purified about 500-fold in 25% yield from the extract of rat liver mitochondria. The enzyme cleaves yeast RNA, poly(U), poly(U), poly(C) and denatured DNA to yield oligonucleotides with 5'-phosphoryl and 3'-hydroxyl ends. The enzyme has a molecular weight of about 60 000, a sedimentation coefficient of 4 S and an isoelectric point of 9.0. The behaviors of RNAase activity of the nuclease are identical with those of DNAase activity in column chromatography as well as in catalytic nature. The affinities of RNAase activity for substrate, Mg2+, spermidine and polyvinyl sulfate are lower than those of DNAase activity. The alkaline nuclease activity measured in the homogenate of regenerating rat liver is not significantly changed.     

Interrelation of the enterotoxigenic properties and the antigenic structure of Escherichia

The enzymatic signs and serological characteristics of Escherichia enterotoxigenic strains isolated from patients with acute intestinal diseases and from healthy persons were studied. The cultures were subdivided into 24 enzymatic variants and classified with 48 serogroups and 61 serovars. The enterotoxigenic properties of the strains were compared with their serological characteristics and enzymatic signs. The strains, isolated from different persons and classified with the same serovar, belonged to the same variant with respect to the type of enterotoxin they produced (only thermostable enterotoxin, only thermolabile enterotoxin, or both), were similar in the degree of their toxigenicity and belonged, as a rule, to the same enzymatic variant. The data on the presence of manifest interrelation between the enteropathogenicity of Escherichia and their structure, as well as on the stability of the enterotoxigenic properties of these organisms, indicate that in acute intestinal diseases the determination of Escherichia enterotoxigenic strains can be carried out by common bacteriological techniques with the use of specific agglutinating sera.     

Intracellular nuclease activities of buffalo rumen bacterial population

Nuclease activities of the predominantly bacterial population obtained from buffalo rumen were investigated. Optimum temperature for hydrolysis of both DNA and RNA was 50°C whereas DNAase activity was observed to be stable up to 50°C, a decrease in RNAase activity was observed even after 40°C. Two pH optima, one at 5.5 and the other at 7.5, were recorded for hydrolysis of DNA. RNAase activity was maximum between pH 6.0 to 7.0. Whereas DNAase activity was stable near its optimum pH, RNAase activity was stable between pH 7.0 to 8.5. Mn²⁺ ions stimulated DNAase activity. It was strongly inhibited by Hg²⁺, Zn²⁺, Pb²⁺ and Ag⁺. RNAase activity was stimulated by Mg²⁺ ions and was strongly inhibited by Hg²⁺, Cu²⁺, Zn²⁺ and Ag⁺. Cysteine hydrochloride and 2-mercaptoethanol stimulated DNAase activity. The activity was strongly inhibited by N-ethylmaleimide, 4-chloromercuribenzoate, 8-quinolinol, iodoacetic acid and 1,10-phenanthroline. RNAase activity was stimulated by cysteine hydrochloride, reduced glutathione and 2-mercaptoethanol and was strongly inhibited by 4-chloromercuribenzoate, 8-quinolinol and 2,2′-bipyridyl. Part of PhD Thesis submitted by the first author to Kurukshetra University.     

The pathogenicity enzymes of clinical strains of Klebsiella pneumoniae]

In 108 K. pneumoniae clinical strains isolated in pneumonia (32 strains), inflammatory processes of the urinary tracts (36 strains) and toxicoseptic states (40 strains) caseinolytic, gelatinase, phosphatase, lecithinase activities and the capacity for producing DNAase and RNAase were studied. The presence of caseinolytic activity was found in 38 cultures (35.1%), gelatinase activity in 37 cultures (34.2%) and lecithinase activity in 13 cultures (12.0%). The production of RNAase was noted in 74 strains (68.5%), DNAase in 56 strains (51.8%) and acidic phosphatase in 33 strains (30.5%). The role of the above-mentioned enzymes in the development of purulent inflammatory processes, as well as the importance of further studies, including those aimed at establishing the nature of the genetic control of the already known properties of the pathogen, are discussed.     

Enterotoxigenic capacity of strains of Klebsiella and Enterobacter genera isolated in acute intestinal diseases in children

234 strains, including 104 K. pneumoniae strains, 28 K. oxytoxica strains, 64 E. cloacae strains and 40 E. aerogenes strains, have been isolated from the intestine of 266 children with diarrhea, aged up to 1 year, and studied for enterotoxigenicity. By the coagglutination test, made with G. Kronvall's staphylococcal reagent prepared with the use of antiserum to Escherichia coli LT-enterotoxin, and the biological assay on suckling mice enterotoxigenic activity has been revealed in 119 strains, including 48 K. pneumoniae strains (12.6%), 33 E. cloacae strains (27.4%) and 23 E. aerogenes strains (19.7%). The strains producing only LT-enterotoxins, only ST-enterotoxins, and both LT- and ST-enterotoxins have been found. The determination of the enterotoxigenic activity of the clinical isolates of opportunistic enterobacteria makes it possible to improve the etiological interpretation of acute intestinal infections.     

甘蔗和烟草幼叶原生质体的核酸含量与核酸酶活性及其影响因素

与幼叶组织相比,酶法新鲜分离的甘蔗和烟草幼叶原生质体内的ＲＮＡ、ＤＮＡ及总核酸含量均降低。其原因可能是刚游离的原生质体内酸性和碱性ＲＮＡ酶与ＤＮＡ酶等活性提高所致。甘蔗叶原生质体内的核酸降低量和ＲＮＡ酶与ＤＮＡ酶活性的增加程度均高于烟草。随用作渗透压稳定剂的甘露醇浓度增加,甘蔗和烟草叶原生质体的ＲＮＡ酶和ＤＮＡ酶活性均相应提高。其中以甘蔗叶原生质体的核酸酶活性增加水平较明显。在细胞壁降解产物的作用下,除了甘蔗原生质体内的ＲＮＡ酶活性略被促进外,其ＤＮＡ酶和烟草叶原生质体内的核酸酶均不受影响     

Isolation and characterization of a ribonuclease III deficient mutant of Escherichia coli

Summary A mutant of E. coli K 12 AB301 RNAase ₁₉ ^- , selected for its inability to degrade double-stranded RNA, has been isolated and shown to have less than 1% of RNAase III-activity related to the parental strain.Abbreviations TCA trichloracetic acid - RF replicative form of phage-RNA Enzymes Lysozyme (E.C. 3.2.1.17) - RNAase (E.C. 2.7.7.16) - DNAase (E.C. 3.1.4.5)     

A rapid high-yield purification procedure for the cyclic adenosine 3',5'-monophosphate receptor protein from Escherichia coli.

A new method for the purification of the cyclic adenosine 3',5'-monophosphate receptor protein from Escherichia coli has been developed. The method is rapid and simple, and a very high yield of the purified protein is obtained which is not contaminated with either DNAase or RNAase activity.     

The putative 15 S precursor of globin mRNA contains a poly (A) sequence

[3H] Uridine or [3H] adenosine pulse-labelled nuclear RNA was isolated from chicken immature red blood cells and separated on denaturing formamide sucrose gradients. RNA of each gradient fraction was hybridized with unlabelled globin DNA complementary to mRNA (cDNA) and subsequently digested by RNAase A and RNAase T1. The experiments revealed two RNA species with globin coding sequences sedimenting 9 S and approx. 15 S, the latter probably representing a precursor of 9 S globin mRNA. A poly (A) sequence was demonstrated in this RNA by two different approaches. Nuclear RNA pulse-labelled with [3H] uridine was fractionated by chromatography on poly (U)-Sepharose. Part of the 15 S precursor was found in the poly(A)-containing RNA. In the second approach 15 S RNA pulse-labelled with [3H]adenosine was hybridized with globin cDNA, incubated with RNAase A and RNAase T1 and subjected to chromatography on hydroxyapatite. The hybrids were isolated and after separation of the strands degraded with DNAase I, RNAase A and RNAase T1. By this procedure poly(A) sequences of approximately 100 nucleotides could be isolated from the 15 S RNA with globin coding sequences. The poly(A) sequence was completely degraded by RNAase T2.     

Changes in the activity of trout lysosomal enzymes during fasting

Activity of lysosome enzymes (acidic phosphatase, beta-glucosidase, DNAase, RNAase and cathepsin D) is determined for its variation in different organs of rainbow trout during complete fasting. It is shown that the activity of most enzymes of concern almost in all organs except skeletal muscles is on the higher level in trouts fasted for 30 days than in the control ones. With an increase of the fasting term to 60 days the acid phosphatase, DNAase, RNAase activity decreases while the glucosidase and cathepsin D activity in some organs increases. Variations detected in the enzyme profile of the trout lysosomes under fasting are of adaptive character.     

A rapid high-yield purification procedure for the cyclic adenosine 3′,5′-monophosphate receptor protein from Escherichia coli

A new method for the purification of the cyclic adenosine 3′,5′-monophosphate receptor protein from Escherichia coli has been developed. The method is rapid and simple, and a very high yield of the purified protein is obtained which is not contaminated with either DNAase or RNAase activity.