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Comparative analysis of the genomes of intestinal spirochetes of human and animal origin.

The aim of the present work was to compare the genomes of 21 strains of intestinal spirochetes, which were isolated from patients suffering intestinal disorders, with those of Treponema hyodysenteriae (strain P18), the known etiological agent of swine dysentery (bloody scours), and of a nonpathogenic strain (M1) of Treponema innocens. The percent guanine-plus-cytosine value of the 23 DNAs was found to be 25.5 to 30.1, as determined by a double-labeling procedure based on nick-translation by DNA polymerase I. The genome size of two spirochetal strains, of human and porcine origin, was found to be similar (4 x 10(6) base pairs) and close to that of the reference bacterium Escherichia coli (4.2 x 10(6) base pairs). Restriction analysis showed the presence of two modified bases in spirochetal DNA. Methyladenine was present in the GATC sequence of DNA from 15 spirochetes of human origin, and methylcytosine was present in several sequences occurring in all strains. The DNA of T. hyodysenteriae displayed a 30 to 100% homology with respect to that of 21 spirochetes from humans, thus suggesting the occurrence of a genetic heterogeneity in the latter group. These data indicate that the intestinal spirochetes analyzed in the present work are related; hence there is a possibility of domestic animals being reservoirs of microorganisms pathogenic for humans. A classification of intestinal treponemes into subgroups has been proposed on the basis of restriction analysis and hybridization experiments.

Animals↗

Effect of addition of a probiotic microorganism to broiler diets contaminated with deoxynivalenol on performance and histological alterations of intestinal villi of broiler chickens.

An experiment was conducted to study the effects of deoxynivalenol (DON) on the performance of broilers, organ weights, and intestinal histology and to evaluate the efficacy of a probiotic feed additive (PB, Eubacterium sp.) with the ability to deepoxidize DON. Two hundred seventy-seven 1-d-old broiler chicks were randomly assigned to 1 of the 3 dietary treatments for 6 wk. The dietary treatments were 1) control; 2) artificially contaminated diets with 10 mg of DON/kg of diet; 3) DON-contaminated diets plus probiotic feed additive (DON-PB). The BW and the efficiency of feed utilization were not adversely affected (P > 0.05) by the inclusion of DON in the diets. A slight improvement in feed intake and BW gain over the course of the experiment was observed in broilers fed DON-PB with no change in feed efficiency. The absolute or relative organ weights were not altered (P > 0.05) in broilers fed the diet containing DON compared with controls and the DON-PB group. The absolute liver weights were numerically increased (P < 0.1) for broilers receiving the diet containing DON-PB. There were no significant differences in the absolute and relative weights of the gizzard, duodenum, pancreas, heart, and spleen. However, the absolute and relative weights of the jejunum and cecum were increased for DON-PB-fed broilers compared with the controls and DON group. No pathological lesions were found in the gut of birds fed DON-contaminated diets during the feeding trial, but mild intestinal changes were observed. The DON altered small intestinal morphology, especially in the duodenum and jejunum, where villi were shorter and thinner (P < 0.05). The addition of the eubacteria to the DON-contaminated feed of the broilers effectively alleviated the histological alterations caused by DON and led to comparable villus length as in the control group. In conclusion, diets with DON contamination below levels that induce a negative impact on health and performance could affect small intestinal morphology in broilers. The histological alterations caused by DON were reduced by supplementing the DON-containing diets with PB. This indicates that in case of DON contamination of feedstuffs, the addition of PB would be a proper way to counteract the possible effects caused by this mycotoxin.

Animals↗

[Intestinal colonization of newborns treated in intensive care units by multiple drug resistant microorganisms].

The aim of the study was to examine the digestive tract colonisation of the newborns by multiple drug resistant bacteria during hospitalization. On the day of admission, after 5 days of hospitalization and at the day of discharge swabs from the anus of the 31 newborns hospitalized in OITiPN were taken and cultured on nutrient and selective media for staphylococci, enterococci, gram negative bacilli and fungi. Susceptibility to antibiotics of bacteria was determined, with giving attention to such resistance mechanisms as: methicillin resistant staphylococci (MRS), high level aminoglycoside resistant (HLAR) and vancomycin resistant enterococci (VRE) and the production of extended spectrum beta-lactamases (type ESBL) by gram negative bacilli. On the day of admission in 7 newborns methicillin resistant staphylococci (MRSCN) were grown in 24 no multiple drug resistant bacteria were found. Among those in 23 already after 5 days of hospitalization, colonization by multiple drug resistant strains was determined: coagulase-negative methicillin resistant staphylococci (MRSCN) were found in 16 children, strains of enterococci (HLAR) in 3 newborns and gram negative ESBL (+) bacilli also in 3 cases. On the day of discharge from hospital (after 13-141 days) in 23 out of 24 newborns enteric tract colonization by multiple drug resistant strains was assessed. In the enteric tract of 3 newborns hospitalized up to 2 weeks coagulase-negative methicillin resistant staphylococci (MRSCN) and/or HLAR enterococci were found; gram negative bacilli that produce ESBL appeared in newborns hospitalized for longer than 14 days. They were isolated in 12 out of 21 newborns. Forming of the enteric tract bacterial flora of the long hospitalized newborns depends on the time of hospitalization as well as on the used therapy.

Drug Resistance, Multiple, Bacterial↗

[Influence of various microbial contents in drinking water on cellular composition of lymphoid plaque in the small intestine of the rat].

Morphological investigations of rats after extended period of drinking water with varying amounts of conventionally pathogenic microflora revealed changes in the cellular composition of different morphological/functional sections of lymphoid plaques in the small intestine which depended on concentrations of microorganisms. Comparing with the control, sites of lymphoid node reproduction were significantly richer in the relative number of plaques and large lymphocytes when water contained 1,000 microbial bodies in 1 cm3. Concentrations of microorganisms higher, the parameters tended down to referential levels. Rise in water concentrations of microorganisms was associated with a decrease in the relative number of small lymphocytes, i.e. main members of the cell in lymphoid nodule mantle with the reproduction site, and an increase in the number of plasmatic cells and macrophages in the diffuse lymphoid tissue. Lymphoid nodes lacking the reproduction sites were observed in the control and in the group of animals who drank minimum microbes (100 cells in 1 cm3). These nodules were viewed again in the rats who had drunk maximum antigens (1 000,000 cells in 1 cm3). These changes point to proliferation and differentiation of lymphoid cells driven by water microorganisms, and inhibition of these processes by water concentrations of conventionally pathogenic microflora above 1,000 microbial bodies in 1 cm3.

Animals↗

Mycobacterium avium interaction with macrophages and intestinal epithelial cells.

Mycobacterium avium is an environmental microorganism that is adapted to live both in the environment (mainly in water and soil) and in bird, fish and mammal hosts. In humans, M. avium infection is seen in patients with some sort of immunosuppression, such as patients with chronic lung disease, and Acquired Immunodeficiency Syndrome. More recently, other populations were shown to be at risk to develop M. avium disease. For the majority of time, humans acquire M. avium through the intestinal tract where the bacterium comes in contact with and translocates the intestinal mucosa. M. avium possesses a unique manner to interact with the intestinal mucosa, and, following invasion, can enter and survive within macrophages and monocytes. Although in vitro entry seems to be dependent on binding to the complement receptor, this finding has not been observed in vivo where the bacterium appears to enter macrophages by alternative mechanisms. The bacterium appears to trigger little inflammatory response, and is able to adapt itself to different environments in the host.

Bacterial Translocation↗

CARD15/NOD2 functions as an antibacterial factor in human intestinal epithelial cells.

BACKGROUND & AIMS: Mutations in the CARD15/NOD2 gene, a putative intracellular pattern recognition receptor, have been linked to the risk for Crohn's disease. Because intestinal epithelial cells play a role as the barrier to luminal microorganisms, we investigated the expression and function of CARD15/NOD2 in intestinal epithelial cells. METHODS: Expression of CARD15/NOD2 messenger RNA (mRNA) in intestinal epithelial cell lines and primary intestinal epithelial cells was assessed by reverse-transcription polymerase chain reaction (RT-PCR). Regulation of expression of CARD15/NOD2 by cytokines was determined by Northern blot using the SW480 cell line. Active CARD15/NOD2 protein in SW480 cells was assessed by the combination of immunoprecipitation and immunoblotting using anti-CARD15/NOD2 antisera. To identify the functional role of CARD15/NOD2 in intestinal epithelial cells, gentamicin protection assays of Salmonella typhimurium were performed using Caco2 cells stably transfected with either wild-type CARD15/NOD2 or the 3020insC mutant associated with Crohn's disease. RESULTS: CARD15/NOD2 mRNA was expressed in both intestinal epithelial cell lines and primary intestinal epithelial cells. CARD15/NOD2 mRNA and protein were up-regulated by tumor necrosis factor alpha (TNFalpha) in SW480 cells. The number of viable internalized S. typhimurium in Caco2 cells stably transfected with CARD15/NOD2 expression plasmid was lower than untransfected Caco2 cells or MOCK transfectant. In contrast, expression of a variant associated with Crohn's disease was unable to constrain bacterial survival. CONCLUSIONS: CARD15/NOD2 is expressed in intestinal epithelial cells and may serve as a key component of innate mucosal responses to luminal bacteria as an antibacterial factor. Failure in this activity may contribute to the development of Crohn's disease.

Caco-2 Cells↗

Manipulation of the oropharyngeal and intestinal microflora by norfloxacin: microbiological and clinical aspects.

During the last few years the impact on the human endogenous microflora by norfloxacin has been studied by several investigators. The use of norfloxacin for selective decontamination in compromised patients and for prophylaxis of recurrent urinary tract infections has also been investigated. This review article summarizes the published data from these studies. The results show that the oropharyngeal flora is only slightly affected by norfloxacin. In the intestinal flora, the Gram-negative aerobic microorganisms are strongly suppressed during administration of norfloxacin. The impact on the Gram-positive aerobic flora is minor, while the anaerobic intestinal microflora is virtually unaffected by the administration. In two studies where norfloxacin was given prophylactically to granulocytopenic patients, the development of aerobic Gram-negative infections was suppressed and overall morbidity decreased. Norfloxacin for long-term prophylaxis of infections in artificially ventilated patients in intensive care units was reported to give encouraging results in one investigation. Two studies indicated that long-term norfloxacin prophylaxis reduces the rate of recurrent urinary tract infections. Norfloxacin administration seems not to predispose for development of resistance to 4-quinolones nor to other commonly used antimicrobial agents.

Bacteria↗

[Method of determining the microbial flora of the small intestinal cavity and mucosa in different gastrointestinal tract diseases].

Intestinal microflora in patients with chronic enterocolitis and duodenal ulcer was studied. Both the cavity of the small intestine and the structures of its mucosa were investigated. The method of aspiration biopsy was used, the samples of multiple washings of the intestinal mucosa and its homogenate were used for seeding. The results thus obtained in indicated that the structures of the mucosa contained microorganisms similar to those isolated from the intestinal cavity, but in lesser amounts and with different characteristics. The proposed method of detecting the microflora in the intestinal cavity and mucosa is simple and can be used in clinical practice.

Adult↗

Use of selective media for detection of cephalothin-resistant bacteria in surgical patients.

Bacterial colonization in hospitalized patients is an important step in nosocomial infections. Frequent employment of antimicrobials can modify patients' normal microbiota, favoring colonization and infection by antimicrobial-resistant microorganisms. First-generation cephalosporins are frequently used as prophylactic antibiotics in surgery. Intestinal, oropharyngeal and skin colonization by cephalothin-resistant microorganisms were studied in 60 pre-operative patients at the Hospital Universitário Pedro Ernesto. Feces were cultured in Eosin-methylene blue medium containing 32 microg/mL of cephalothin. Swabs obtained from the oropharynx and from skin were inoculated in cistein-lactose electrolytes-deficient medium containing 32 microg/mL of cephalothin. Isolated strains were identified and tested for susceptibility to antimicrobials by disk diffusion. Cephalothin-resistant strains were isolated from the feces of 59 patients (98%), from the oropharynx of 13 patients (22%) and from skin in 10 patients (17%). Enterobacter cloacae was predominant in feces (68% of the patients) and oropharynx (13%). Acinetobacter spp. was the most frequent microorganism isolated from the skin (10%). Antimicrobial multiresistant strains were isolated from at least one of the sites in 38 patients (63%). The employment of selective medium containing antimicrobials is a relatively simple and efficient method, being useful to evaluate microorganisms from hospitalized patients' microbiota that are relevant as potential pathogens in nosocomial infections.

Anti-Bacterial Agents↗

Alterations in intestinal transport of oxalate in disease states.

Normally, absorption of oxalate from dietary sources can occur in all segments of the intestinal tract. However, alterations in both the magnitude and direction of oxalate fluxes across the intestine can occur in disease states. In enteric hyperoxaluria, enhanced absorption of oxalate by the large intestine is caused by increased permeability of a shunt conductance induced by malabsorbed bile salts and fatty acids. In this condition, the contribution of a paracellular passive flux of oxalate moving along its electrochemical gradient will predominate when intraluminal concentrations of free oxalate are high. In contrast, in chronic renal failure, secretion of oxalate can occur across both small and large intestine thereby facilitating extrarenal elimination with subsequent degradation by mucosal substrate-specific microorganisms. Clearly, in recent studies of oxalate transport, the intestine has emerged with an integral role in mass balance of oxalate in health and disease.

Animals↗

[Factors in the persistence of opportunistic microorganisms in dysbacteriosis of the gastrointestinal tract].

The presence of correlation between the quantitative content of individual carboxylic acids and aromatic compounds of microbial origin in feces and the degree of the antagonistic activity of feces was proved in experiments on conventional laboratory rats receiving therapeutic doses of tetracyclin and pefloxacin for 8 days. Antagonistic activity was regarded as one of the factors of the colonization resistance of the intestine to the invasion of opportunistic microorganisms, and microbial metabolites studied in this work were regarded as factors contributing to the persistence of these microorganisms.

4-Quinolones↗

Intestinal mucins in colonization and host defense against pathogens.

Intestinal mucins are key components of the first line of host defense against intestinal pathogens. These large glycoconjugates secreted by specialized exocrine goblet cells form viscous gels that trap microorganisms and irritants and limit their diffusion to the intestinal epithelium. Moreover, they allow for colonization by indigenous bacterial flora that prevents attachment of pathogenic microbes. The interaction between microbes and mucins involves mucin carbohydrate side chains and microbial adhesin molecules. Certain microorganisms and disease states may alter mucin biochemistry or expression. Although these alterations most likely contribute to disease processes, the full impact of these phenomena are still unclear. The development of mucin-secreting cell lines has facilitated the study of mucin biology and aided our understanding of mucin-microbial interactions.

Animals↗

[Microflora study of the intestinal cavity and mucosa normally and after gamma irradiation].

Lactobacilli and bifidobacteria are numerically prevalent in the microflora of the mucous membrane in mice and guinea-pigs. Some opportunistic microorganisms (Proteus, enterococci, aerobic bacilli, clostridia in mice; streptococci, yeast, enterococci in guinea-pigs) can mainly be found in the cavity of the large intestine, only small amounts of these microorganisms (10(2)), if any, are present on the mucous membrane. In the course of postradiation dysbacteriosis in mice and guinea pigs all opportunistic microorganisms which inhabit the cavity of the large intestine, proliferating actively, disseminate into the small intestine and, probably, invade the mucous membrane. Such microorganism as Escherichia, Proteus, Clostridium, normally absent in intact animals, appear in the intestine of guinea of guinea pigs subjected to irradiation. At the same time lactobacilli and bifidobacteria sharply decrease in number.

Animals↗

Clostridial enteric infections in pigs.

Clostridium perfringens types A and C and Clostridium difficile are the principal enteric clostridial pathogens of swine. History, clinical signs of disease, and gross and microscopic findings form the basis for a presumptive diagnosis of C. perfringens type-C enteritis. Confirmation is based on isolation of large numbers of type-C C. perfringens and/or detection of beta toxin in intestinal contents. Diagnosis of C. perfringens type-A infection, however, remains controversial, mostly because the condition has not been well defined and because type-A organisms and their most important major (alpha) toxin can be found in intestinal contents of healthy and diseased pigs. Isolation of large numbers of C. perfringens type A from intestinal contents, in the absence of other enteric pathogens, is the most reliable criterion on which to base a diagnosis. Recently, beta2 (CPB2) toxin-producing C. perfringens type A has been linked to disease in piglets and other animals. However, implication of CPB2 in pathogenesis of porcine infections is based principally on isolation of C. perfringens carrying cpb2, the gene encoding CPB2, and the specific role of CPB2 in enteric disease of pigs remains to be fully defined. Clostridium difficile can also be a normal inhabitant of the intestine of healthy pigs, and diagnosis of enteric infection with this microorganism is based on detection of its toxins in feces or intestinal contents.

Animals↗

Inhibition of Streptococcus mutans NS adhesion to glass with and without a salivary conditioning film by biosurfactant- releasing Streptococcus mitis strains.

The release of biosurfactants by adhering microorganisms as a defense mechanism against other colonizing strains on the same substratum surface has been described previously for probiotic bacteria in the urogenital tract, the intestines, and the oropharynx but not for microorganisms in the oral cavity. Two Streptococcus mitis strains (BA and BMS) released maximal amounts of biosurfactants when they were grown in the presence of sucrose and were harvested in the early stationary phase. The S. mitis biosurfactants reduced the surface tensions of aqueous solutions to about 30 to 40 mJ m(-2). Biochemical and physicochemical analyses revealed that the biosurfactants released were glycolipids. An acid-precipitated fraction was extremely surfactive and was identified as a rhamnolipidlike compound. In a parallel-plate flow chamber, the number of Streptococcus mutans NS cells adhering to glass with and without a salivary conditioning film in the presence of biosurfactant-releasing S. mitis BA and BMS (surface coverage, 1 to 4%) was significantly reduced compared with the number of S. mutans NS cells adhering to glass in the absence of S. mitis. S. mutans NS adhesion in the presence of non-biosurfactant-releasing S. mitis BA and BMS was not reduced at all. In addition, preadsorption of isolated S. mitis biosurfactants to glass drastically reduced the adhesion of S. mutans NS cells and the strength of their bonds to glass, as shown by the increased percentage of S. mutans NS cells detached by the passage of air bubbles through the flow chamber. Preadsorption of the acid-precipitated fraction inhibited S. mutans adhesion up to 80% in a dose-responsive manner. These observations indicate that S. mitis plays a protective role in the oral cavity and protects against colonization of saliva-coated surfaces by cariogenic S. mutans.

Bacterial Adhesion↗

Microbial colonization of the intestinal epithelium in suckling mice.

Colonization by indigenous microorganisms of the mucosal epithelia of the large bowels of suckling mice was followed by microbial culture techniques and by light, fluorescence, and electron microscopy. Certain microbes colonize in distinctive patterns the cecal and colonic epithelia in these mice. Coliforms and enterococci colonize the large bowel 7 to 9 days after birth and reach high population levels during the second week. During that period, these facultative anaerobes can be detected by immunofluorescence techniques in microcolonies in the mucin on the epithelium. During the third week, however, after their populations decline to the low levels characteristic of adult mice, coliforms and enteroccoci can be observed only infrequently in the mucous layer. Anaerobic fusiform-shaped bacteria appear in the mucous layers along with the microcolonies of enterococci and coliforms during the second week after birth. These anaerobes increase in numbers in the mucin until they form thick layers on the mucosal epithelium by the end of the third week. They remain in the mucous layer throughout the life of the normal mouse. Anaerobic spiral-shaped microbes also colonize the mucous layer on the cecal and colonic epithelium. But these organisms can be detected by immunofluorescence in 1-week-old mice, well in advance of the time the fusiform-shaped bacteria can be found. In the second week, the latter microbes co-inhabit the mucous layer with the spiral-shaped organisms. The fusiform- and spiral-shaped microbes remain associated in the mucin on the cecal and colonic mucosal epithelia into the adult life of mice.

Animals↗

[Lactose--a potential dietary fiber. The regulation of its microecological effect in the intestinal tract. 2. The nutrient effect of lactose].

In the small intestine lactose is subjected to the hydrolytic impact of beta-galactosidase originating mainly from the mucosa. In rats about two thirds of the enzyme activity are located in the first part of the small intestine, and one third in the second one. A part of the mucosal enzyme does not remain in the mucosa. It becomes detached and can be determined in the chymus. The ratio of the transient to the resident proportion amounts to 1.8: 1 in germfree and 0.23: 1 in conventional rats. Bacterial settlement causes an increase in the mucosal mass resulting in higher total activity whereas the specific activity of the mucosal enzyme remains unchanged. Microorganisms occurring close to the small intestine mucosa take part in lactose degradation. Lactose-containing diet leads to an increase in both the bacterial and the mucosal activity, the latter one to varying degrees. Lactose concentration in the ileal chymus rises with increasing intake of lactose and decreasing protein and phosphate intake. Following a saturation kinetics the velocity of lactose hydrolysis is correlated with the lactose concentration of the diet. alpha-lactose is hydrolysed more rapidly in the small intestine of both human sucklings and rats than beta-lactose. As the results of a mathematical model show lactose mutarotation does not effect on the degree of lactose degradation in the small intestine. Depending on the intake of lactose and the food composition the rate of lactose hydrolysis amounts to: --max. 50% after small intestine perfusion in human sucklings, --max. 80% after small intestine perfusion in rats, --max. 60% in rats with ileostomata.

Animals↗

External decontamination of wild leeches with hypochloric acid.

BACKGROUND: Medicinal leech, Hirudo medicinalis, has been used in plastic and reconstructive surgery, to relieve venous congestion and to improve the microrevascularization of flaps. In many countries, wild leeches are still provided from local markets and utilised with antibiotic prophylaxies. In this research, results of identification of bacteria in the transport fluid is reported, oral and intestinal floras and the antibiograms of the identified microorganisms are investigated. Also, to avoid possible infections, the ability of hypochloric acid, a disinfectant, to suppress the relevant microorganisms without changing the life style and behavior of leeches in terms of sucking function, is investigated. METHODS: Bacterial identifications and antibiograms of oral and intestinal flora and transport medium were performed for 10 leeches. The optimum concentration of hypochloric acid which eliminated microorganisms without affecting the viability and sucking function of the leeches were determined by dilution of hypochloric acid to 100, 50, 25, 12.5, 6.25 ppm concentrations in different groups of 25 leeches. Finally, 20 leeches were applied atraumatically to the bleeding areas of rats, the duration of suction was determined and compared statistically between the leeches treated and not treated with hypochloric acid solution. RESULTS: Aeromonas hydrophilia was the most commonly identified microorganism and found to be resistant to first generation cephalosporins, frequently used in prophylaxis at surgical wards. In the next stages of the study, the leeches were subjected to a series of diluted hypochloric acid solutions. Although disinfection of the transport material and suppression of the oral flora of hirudo medicinalis were successful in 100, 50, 25, 12.5, 6.25 ppm concentrations; 12.5 ppm solution was the greatest concentration in which hirudo medicinalis could survive and sucking function was not affected significantly. CONCLUSIONS: External decontamination of wild leeches with 12.5 ppm hypochloric acid enables bacterial suppression without causing negative effects on leech sucking function and life.

Aeromonas↗