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[Use of lincomycin, methicillin and ristomycin in the nutrient media for isolating pathogenic intestinal microorganisms].

Elective-differentiating solid nutrient media for simultaneous isolation of Vibrioes, Salmonella and Shigella were developed. Antibiotics active against grampostive microflora and dry bile salts inhibiting the growth of Proteus were used as the inhibitors of the growth of the accompanying microflora. The medium was lincomycin and the bile salts may be prepared in a dry form.

Acute Disease↗

Further studies on the hydrolysis of salicyluric acid in intestinal microorganisms and prolonged blood concentration of salicylic acid following rectal administration of salicyluric acid in rabbits.

The blood concentrations of salicyluric acid and salicylic acid following intracecal and rectal administration of salicyluric acid were determined in rabbits. Immediate and very extensive salicylic acid formation in the cecum was found following intracecal administration. After rectal administration, a small amount of salicyluric acid was absorbed in intact form. The rest was rapidly hydrolyzed to salicylic acid, which was subsequently absorbed. The blood concentration of salicylic acid was maintained at 1.3-1.8 micrograms/ml from 2 to 12 h. Three doses of salicyluric acid were administered rectally. The peak level of salicyluric acid increased with dose. However, salicylic acid concentration in the blood following administration of salicyluric acid at 10.0 mg/kg (salicylic acid equivalent) was not double that observed following administration of salicyluric acid at 5.0 mg/kg (salicylic acid equivalent). It appears that a larger amount of salicyluric acid in the rectal lumen may have saturated the glycine deconjugation system.

Administration, Rectal↗

Hydrolysis of salicyluric acid in intestinal microorganisms and prolonged blood concentration of salicylic acid following rectal administration of salicyluric acid in rats.

The blood concentrations of salicyluric acid and salicylic acid following oral, intravenous, intracecal and rectal administration of salicyluric acid were determined in rats. After oral administration of salicyluric acid, salicyluric acid was rapidly absorbed. Salicylic acid was detected at low concentration. Following intravenous administration of salicyluric acid, salicyluric acid was detected in the blood and was rapidly eliminated. A trace amount of salicylic acid was detected, suggesting that systemic deconjugation of glycine was involved. Furthermore, in vitro incubation of salicyluric acid with contents of the gut showed that the major source of the hydrolysis was the hind gut. Immediate and very extensive salicylic acid formation in the cecum was found following intracecal administration of salicyluric acid. The blood concentration of salicylic acid was maintained at 2.6-4.0 micrograms/ml from 4 to 12 h following rectal administration of salicyluric acid (10 mg/kg: salicylic acid equivalent). Species difference in the metabolic fate of salicyluric acid in rats and rabbits reported previously is discussed.

Administration, Oral↗

Isolation and characterization of thirteen intestinal microorganisms capable of 7 alpha-dehydroxylating bile acids.

Thirteen anaerobic bacteria capable of performing the 7 alpha-dehydroxylation of both cholic acid and chenodeoxycholic acid were isolated from human feces and also from sewage. Ten organisms from heat-treated samples were species of Clostridium identical or closely related to the Clostridium bifermentans-C. sordellii group and consisted of four strains elaborating 7 alpha-dehydroxylase alone and six strains capable of catalyzing both 7 alpha-dehydrogenation and 7 alpha-dehydroxylation. The remaining three organisms, recovered from fresh human feces, were gram-positive, nonflagellated, nonsporeforming, anaerobic rods and comprised two distinct species. Strain HD-17, still unidentified, had both activities, but was unique in that it exclusively 7 alpha-dehydroxylated cholic acid while biotransforming chenodeoxycholic acid, preferably though 7 alpha-dehydrogenation. Two unclassified strains, b-8 and c-25, metabolized both acids though 7 alpha-dehydroxylation and 7 alpha-dehydrogenation. Except for strains b-8 and c-25, all of th 7 alpha-dehydroxylating bacteria split the conjugated bile acid series, and hydrolases were detected in cell-free filtrates of early stationary-phase broth cultures.

Bacteria↗

Cooperative formation of omega-muricholic acid by intestinal microorganisms.

Three anaerobic bacteria, isolated from the ceca of rats and mice, converted, through a concerted mechanism, beta-muricholic acid, the predominant bile acid in germfree rats, into omega-muricholic acid. One isolate was a Eubacterium lentum strain; the second and third isolates were tentatively identified as atypical Fusobacterium sp. strains. The conversion of beta-muricholic acid into omega-muricholic acid proceeded in two steps: E. lentum oxidized the 6 beta-hydroxyl group of beta-muricholic acid to a 6-oxo group, which was reduced by either of the two other species to a 6 alpha-hydroxyl group, yielding omega-muricholic acid. This transformation occurred both in vitro and in gnotobiotic rats. Monoassociation of germfree rats with the E. lentum strain gave rise to an unidentified fecal bile acid, probably a derivative of beta-muricholic acid having a double bond in the side chain.

Animals↗

Conversion of 7-ketolithocholic acid to ursodeoxycholic acid by human intestinal anaerobic microorganisms: interchangeability of chenodeoxycholic acid and ursodeoxycholic acid.

Chenodeoxycholic acid, ursodeoxycholic acid and 7-ketolithocholic acid were incubated with human intestinal bacteria (source: 4 healthy males) at 37 degrees C for 72 hours in an anerobic condition. The bile acids of the products in culture medium were identified by three independent methods, thin layer chromatography, gas-liquid chromatography and GLC-mass spectrometry. Lithocholic acid, ursodeoxycholic acid and 7-ketolithocholic acid were observed in the culture of chenodeoxycholic acid. Lithocholic acid, chenodeoxycholic acid and 7-ketolithocholic acid were observed in the culture of ursodeoxycholic acid. Chenodeoxycholic acid and ursodeoxycholic acid were produced from 7-ketolithocholic acid. These data may suggest that chenodeoxycholic acid and ursodeoxycholic acid are interconvertible via 7-ketolithocholic acid by the mixed culture of human intestinal microorganisms under an anaerobic condition.

Anaerobiosis↗

Animal health and foodborne pathogens: enterohaemorrhagic O157:H7 strains and other pathogenic Escherichia coli virotypes (EPEC, ETEC, EIEC, EHEC).

The majority of interactions between microorganisms and animals are based on convenient relations for both of them. Symbiotic microorganisms, like intestinal microbiota, produce important vitamins for animals and protects them from putative pathogens. In general, for monogastric animals, the main contribution of intestinal microorganisms is to supply with growth factors the animal diet, and in some cases they are responsible for providing essential vitamins (e.g. vitamin K). Some particular and relatively few microbes like viruses, bacteria, fungi, protozoa and algae are responsible for animal illness. Because microorganisms are easily dispersed, display physiological diversity, and tolerate extreme conditions, they are ubiquitous and may contaminate and grow in many products, including food and raw materials. Foodborne diseases are caused by consumption of contaminated food or beverages. Many different disease-causing pathogens can contaminate food, so there are many different foodborne infections. In addition, poisonous chemicals and biological toxins can cause disease if they are present in food. To know how a particular disease is spreading is an important matter to take appropriate steps to stop it. For example Escherichia coli O157:H7 infections can spread through contaminated food (meat, vegetables, cheese, etc.), contaminated drinking water or juices, contaminated swimming water and from person to person. Among foodborne pathogens, the most frequently detected are bacteria, but also parasitic protozoa and worms, viruses, natural toxins and other pathogenic agents like prions are important agents for foodborne diseases. Particular pathogenic types of E. coli, classified by their specific pathogenic mechanisms (toxins, adhesins, invasiveness, etc.) are actually known as E. coli virotypes. Enterohaemorrhagic E. coli (EHEC), which constitute the main part of this review, were also named verotoxigenic E. coli (VTEC) or Shiga toxigenic E. coli (STEC). EHEC strains cause haemorrhagic colitis (HC), haemolytic uremic syndrome (HUS) and thrombotic thrombocytopaenic purpura (TP) in humans. They synthetize shigatoxins (verotoxins) which are potent cytotoxic substances, adherence factors and enterohaemolysin. EHEC are responsible for many outbreaks of bloody diarrhoea caused by contaminated foods: beef, milk, fruits, juice, water, etc. The most important serogroups among EHEC are O26, O111 and O157, being O157:H7 the most relevant serotype in foodborne outbreaks. The normal intestinal microflora of cattle was found to be the most relevant reservoir of EHEC strains.

Animals↗

[Phylogenetic analysis of intestinal bacterium using the distribution of intestinal bacterium in cyprinidate fishes and 16S DNA sequence data set].

There is a complex- and multi-effect for interdependent survival between intestinal- microorganisms and hosts. The symbiosis or coevolution that results from this effect for interdependent survival is used to reveal the phylogenies of hosts as well as intestinal microorganisms. The symbiosis or coevolution between intestinal microorganisms and hosts has been generated by interactive natural selection occurred between them. The symbiosis information that has been formed by interactive natural selection during a long evolutionary process must be recorded in DNA sequences. According to this point of view,we analyzed the phylogeny of 9 intestinal bacteria genera using their contents in intestines of 8 Cyrinidate species. At the same time,we fetched the 16S rRNA gene DNA sequences of 43 intestinal bacteria species being included in these nine genera of six intestinal families from GeneBank and constructed phylogenetic trees by NJ and MP methods. The NJ tree and MP tree have the same topologic configuration and are identical with the classical phylogenetic tree. Both the trees of 16S rRNA gene separated 43 bacteria species into gram-negative bacteria group and the gram-positive bacteria group,which are the first branches. Each of the first branches (groups) made again 6 subbranches (subgroups) where each subbranch is a family.Especially,the subbranch (subgroup) of enterobacteriaceace made again four small branches as genus taxon. This tree also shows that bacilliform bacterium is distinct from each other in the NJ and MP trees. After all species on the tree are merged,the topological configuration of the unrooted tree of 16S gene is closed to that of the host range unrooted tree.However,the position of bacillus is greatly changed on both the unrooted trees. The difference can be found if we increase the examination level and extend the hosts examed.

English Abstract↗

Metabolism of vitamin K and influence on prothrombin time in milk-fed preruminant calves.

The metabolism of vitamin K was studied in 66 preruminant veal calves that were fed supplemental menadione sodium bisulfite complex or phylloquinone. Menadione sodium bisulfite complex was converted by intestinal microorganisms to menaquinone-4 and absorbed and stored in the liver as menaquinone-4. Phylloquinone was absorbed unchanged. Production of menaquinones 6, 7, 8, and 10 by intestinal microorganisms also was observed, but was not dependent upon dietary vitamin K. No difference was noted in prothrombin time among the groups. Intestinal microorganisms provide sufficient vitamin K to meet the physiological needs of calves fed milk replacers. Menaquinone-4 was the form of vitamin K used to meet the calf's requirement.

Absorption↗

Where do the immunostimulatory effects of oral proteolytic enzymes ('systemic enzyme therapy') come from? Microbial proteolysis as a possible starting point.

Enteric-coated proteolytic enzyme preparations like Wobenzym and Phlogenzym are widely used for the so-called 'systemic enzyme therapy' both in humans and animals. Numerous publications reveal that oral proteolytic enzymes are able to stimulate directly the activity of immune competent cells as well as to increase efficiency of some of their products. But origins of the immunostimulatory effects of oral proteolytic enzymes are still unclear. The hypothesis described here suggests that it may be proteolysis of intestinal microorganisms that makes the immune competent cells to work in the immunostimulatory manner. The hypothesis was largely formed by several scientific observations: First, microbial lysis products (lipopolysaccharides, muropeptides and other peptidoglycan fragments, beta-glucans, etc.) are well known for their immunostimulatory action. Second, a normal human being hosts a mass of intestinal microorganisms equivalent to about 1 kg. The biomass (mainly due to naturally occurring autolysis) continuously supplies the host's organism with immunostimulatory microbial cell components. Third, the immunostimulatory effects resulting from the oral application of exogenously acting antimicrobial (lytic) enzyme preparations, such as lysozyme and lysosubtilin, are likely to be a result of the action of microbial lysis products. Fourth, cell walls of most microorganisms contain a considerable amount of proteins/peptides, a possible target for exogenous proteolytic enzymes. In fact, several authors have already shown that a number of proteases possess an ability to lyse the microbial cells in vitro. Fifth, the pretreatment of microbial cells (at least of some species) in vitro with proteolytic enzymes makes them more sensitive to the lytic action of lysozyme and, otherwise, pretreatment with lysozyme makes them more susceptible to proteolytic degradation. Sixth, exogenous proteases, when in the intestines, may participate in final steps of food-protein digestion. The resulting food-borne peptides have recently been shown to be potential activators of microbial autolysis. The main question that needs to be answered in order to verify the hypothesis is whether oral proteases are able (and to what extent) to lyse/mediate lysis of intestinal microorganisms in situ. Methods based on up-to-date molecular biology techniques to allow investigation of the influence of exogenous proteases on microbial lysis processes in vivo (in the intestines) need to be developed. Research testing of this hypothesis may have an important impact in development of novel preparations for the systemic enzyme therapy.

Adjuvants, Immunologic↗

[Association of Candida albicans fungi with some opportunistic microorganisms in intestinal dysbiosis in patients of different age groups].

The occurrence of C. albicans in association with opportunistic microorganisms (Staphylococcus aureus and Klebsiella) in intestinal dysbiosis in patents of different age groups (from 6 days to 31 years) was analyzed. The data on the comparative evaluation of the results of the bacteriological examination of patients with intestinal dysbiosis, carried out during the periods of 2000-2001 and 2001-2002 (388 and 467 patients respectively), are presented. During the period of 2000-2001 the detection rate of C. albicans in monoculture was 6-33% of cases (in the examined group). A higher detection rate was registered with respect to the association of C. albicans with staphylococci (42-65% of cases). The associations of C. albicans with staphylococci and Klebsiella were observed in 13-46% of cases. Similar results were registered during the period of 2001-2002, but during this period a reliable decrease in the detection rate of the association of C. albicans with staphylococci (practically by 20% in the group of infants) and the 1 1/2-fold increase of the detection rate of C. albicans in monooulture (in all age groups) were observed.

Adult↗