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Failure of macrophage activation in experimental obstructive jaundice: association with bacterial translocation.

Bacterial translocation from the gastrointestinal tract and macrophage activation are central to current theories of sepsis. The relevance of both in obstructive jaundice is unclear. The effect of bile duct ligation for 7 days on bacterial translocation to mesenteric lymph nodes and on macrophage activation in a rat model was examined. Compared with an incidence of zero in sham-ligated controls, bile-duct ligated rats had a 67 per cent incidence of Gram-negative colonization of mesenteric lymph nodes. This was associated with a significant (P < 0.001) decrease in macrophage tumour necrosis factor, superoxide anion and nitric oxide production compared with that in sham controls. Spontaneous bacterial translocation occurs in experimental obstructive jaundice and is associated with marked suppression of macrophage activation. This suggests a mechanism whereby jaundiced patients may be more susceptible to persistent infection but relatively protected against uncontrolled sepsis.

Animals

Bacterial translocation is bacterial species dependent: results using the human Caco-2 intestinal cell line.

BACKGROUND: Because of in vivo limitations, the mechanisms underlying the process of bacterial translocation are poorly understood. Thus, an in vitro model system to study the translocation process was developed. METHODS: Transformed human colonic carcinoma (Caco-2) intestinal cells were grown as a polarized monolayer on semi-permeable membranes contained in the upper compartment of a two-compartment system. Once the Caco-2 monolayer had reached confluence, the ability of six different species of bacteria to translocate across the monolayer was tested using a dose-response curve of bacteria (10(2), 10(4), or 10(6)) organisms. RESULTS: At inocula of 10(4) or 10(6) but not 10(2) organisms, bacteria crossed the monolayer. Bacterial passage across the monolayer was bacterial species specific, with gram-negative enterics being the best, gram-positive aerobes being intermediate translocators, and strict anaerobes being the poorest translocators. CONCLUSION: The results of this study indicate that bacteria translocate across the Caco-2 monolayer in a dose-dependent and species-related fashion and support the use of this in vitro epithelial cell culture system as a model for studying bacterial transport.

Bacterial Physiological Phenomena

Influence of malnutrition on the prevalence of bacterial translocation and spontaneous bacterial peritonitis in experimental cirrhosis in rats.

Bacterial translocation (BT) has been involved in the pathogenesis of spontaneous bacterial peritonitis (SBP) in experimental cirrhosis. Because malnutrition is a common feature in cirrhosis, the aim of this study was to evaluate the effect of nutrition on BT and SBP. We induced cirrhosis in 44 Sprague-Dawley rats by administration of oral CCl4, and, afterward, 26 animals were maintained with dietary restriction. Cultures of mesenteric lymph nodes (MLN), peripheral and portal blood, liver, and spleen were performed. SBP occurred in 48% of the rats with ascites, this being more frequent in the malnourished animals (80%) than in control rats (29%). BT appeared in all the rats with SBP (100%) but only in 57% without it. In the malnourished animals, the BT rate was 95%, while it was 30% in the control group. These results suggest that malnutrition increases the BT rate and the risk of developing SBP in experimental cirrhosis, and that BT is frequent in cirrhosis and may play a role in the development of SBP.

Animals

[Development of spontaneous bacterial peritonitis in an experimental model in cirrhotic rats. Relationship with intestinal bacterial translocation].

Spontaneous bacterial peritonitis (SBP) is one of the most important complication in cirrhotic patients with ascites, but is pathogenesis is not well known. It is thought that the impaired host defences and the passage of enteric bacteria into the mesenteric lymph nodes, named bacterial translocation, may be two important mechanisms in the pathogenesis of SBP. We have studied this phenomenon in an experimental model with oral CC14 induced cirrhotic rats. SBP occurred in 36% of ascitic rats, all cases being produced by enteric Gram (-) bacteria. Bacterial translocation was observed in 100% of rats with SBP but in 53% of rats without SBP (p < 0.05). In all cases the same organism was isolated in ascitic fluid and in mesenteric lymph nodes. These results suggest that bacterial translocation could play an important role in the pathogenesis of SBP.

Animals

Effects of the angiotensin converting enzyme inhibitor enalapril on bacterial translocation after thermal injury and bacterial challenge.

Burn injury and sepsis produce acute gastrointestinal derangements that may predispose patients to bacterial translocation. We studied the effects of enalapril, an angiotensin converting enzyme inhibitor (ACEI), on gastrointestinal anatomic alterations, bacterial translocation, and related mortality during gut-derived sepsis in burned mice that had received a prior bacterial challenge. BALB/c mice (n = 111) were treated with enalapril 10 or 1 mg/kg body weight or sterile saline as control twice daily for 3 days. They were then gavaged with 10(a)111 in radiolabeled or unlabeled Escherichia coli and given a 20% total body surface area (TBSA) burn injury. Animals gavaged with unlabeled bacteria were observed for survival (n = 60). Survival was significantly higher in the group receiving enalapril 10 mg/Kg compared with control (75% vs. 10%). Mice treated with enalapril maintained small intestine weight, measured 4 h postburn, and ileal mucosal height was preserved, whereas burned untreated animals lost intestinal weight and mucosal height. Bacterial translocation was decreased in mice treated with enalapril, but killing was unaffected. This study suggests that treatment with enalapril positively affects the outcome in gut-derived sepsis by ameliorating gastrointestinal structural and functional damage and decreasing bacterial translocation.

Angiotensin-Converting Enzyme Inhibitors

Effect of alcohol on bacterial translocation in rats.

Bacterial translocation has been proposed to be important in the pathophysiology of sepsis, as well as to be a consequence of sepsis. To study the effect of alcohol on bacterial translocation from the gut, normal Sprague-Dawley rats were administered alcohol by gavage by two regimens: Acute (3.7 g/kg, one dose) or Subacute (1 of 2 doses, 2.4 or 3.7 g/kg/day once daily for 14 days). Mesenteric lymph node cultures were performed, and portal venous blood was assayed for endotoxin. Ileal and cecal permeability studies were performed in the Acute and Subacute groups using fluorescein isothiocyanate-labeled dextrans of either 4,000 or 70,000 kDa size. As an index of the effect of systemic endotoxin, tissues from mesenteric lymph nodes, liver, and intestinal Peyer's patches were assayed for the presence of mRNA for tumor necrosis factor. Additionally, because extrapulmonary sepsis has been shown to suppress pulmonary antibacterial defenses, animals in the Subacute group were challenged by aerosol inoculation with Pseudomonas aeruginosa to determine bacterial clearance and alveolar cellular responses. The results show that neither of the alcohol regimens resulted in bacterial growth from mesenteric lymph nodes or portal blood. Animals in the Subacute group had more endotoxin present in portal blood than did the Control group (92.9 pg/ml vs. 40.2 pg/ml; p < 0.02). None of the animals had demonstrable mRNA for tumor necrosis factor in any of the tissues assayed. There were no demonstrable increases in ileal or cecal permeability for either the small or large molecular weight dextran in either alcohol group. Furthermore, there was no delay in the clearance of P. aeruginosa from the lung in the Subacute group, but these animals recruited fewer neutrophils into the airspaces in response to this challenge than did the Control animals. Thus, alcohol intoxication does not result in bacterial translocation from the gut in this model. Despite higher levels of portal venous endotoxin in the animals in the Subacute alcohol group, no adverse systemic consequences of this phenomenon could be demonstrated.

Alcohol Drinking

Bacterial translocation in the neonate.

Bacterial translocation has been a major topic of investigation for the past two decades. Despite recent evidence that bacterial translocation may play a significant role in the morbidity and mortality of adults faced with multiple types of stress, very little is known about the effect of bacterial translocation on the neonate. Recently more and more evidence has suggested that normal as well as premature or ill neonates experience spontaneous bacterial translocation quite commonly. This article reviews the recent literature on bacterial translocation in stressed adults, the development of an intact intestinal mucosal barrier in the newborn as a protection against bacterial translocation, and the role of spontaneous bacterial translocation in the development of systemic sepsis and its accompanying morbidity and mortality.

Adult

Nitric oxide-derived urinary nitrate as a marker of intestinal bacterial translocation in rats.

BACKGROUND/AIMS: Bacterial translocation across the gut wall may lead to bacteremia and sepsis. Bacteriological analyses are laborious and time consuming, which precludes a rapid diagnosis of bacterial translocation. Synthesis of nitric oxide by macrophages is a primary response to bacterial infections. Therefore, the aim of this study was to examine whether NO-derived nitrate excretion in urine can be used as a rapid and quantitative marker of intestinal bacterial translocation. METHODS: The kinetics of urinary nitrate excretion was determined in rats intraperitoneally injected with increasing doses of Salmonella enteritidis lipopolysaccharide. Subsequently, the response to bacterial translocation was studied in rats infected orally with different doses of viable, invasive S. enteritidis. RESULTS: Increasing the lipopolysaccharide dose from 0.05 to 0.50 mg/kg resulted in a transient, dose-dependent, almost 10-fold increase in urinary nitrate excretion. Administration of the NO synthase inhibitor NG-nitro-L-arginine methyl ester merely inhibited the increase in nitrate excretion after lipopolysaccharide injection. Increasing the infective dose of viable Salmonella resulted in a time- and dose-dependent exponential increase in nitrate output. Translocation was a prerequisite for provoking a nitrate response. Total urinary nitrate excretion after infection and classical infection parameters, such as weight of the mesenteric lymph nodes and population levels of Salmonella in feces, were highly correlated. CONCLUSIONS: Urinary nitrate excretion is a quantitative, noninvasive biomarker of intestinal bacterial translocation, which can be used to follow the course of a systemic infection.

Animals

The effect of surgical trauma on the bacterial translocation from the gut.

Bacterial translocation is the passage of viable bacteria from the lumen of the gastrointestinal tract through the intestinal mucosa to other sites. It is believed that bacterial translocation may lead to infection and septicemia. The purpose of this study was to determine what factors in experimental surgical trauma lead to bacterial translocation. Two-month-old Wistar albino rats were divided into five groups: (A) control; (B) anesthesia (ether inhalation); (C) anesthesia and surgery (median laparotomy and transient compression of the intestines); (D) fasting only; and (E) anesthesia, surgery, and fasting. After 48 hours, ileum, mesenteric lymph nodes, and blood were cultured for aerobic and anaerobic organisms. In each group the number of animals with bacteria overgrowth was calculated. The incidence of bacterial translocation to mesenteric lymph nodes and blood in groups B and D were similar to the controls (P greater than .01). There was a significant increase in the number of animals with bacterial translocation in groups C and E (P less than .001). The majority of translocating bacteria were E coli.

Animals

[Bacterial translocation in Crohn disease].

Bacterial translocation is the passage of viable endogenous bacteria from the gastrointestinal tract to mesenteric lymph nodes and other internal organs. The aim of this work was to study bacterial translocation in patients operated on for Crohn's disease. Twenty-eight patients, mean age 29 years, not having received any antibiotics since at least 8 days, presenting with ileal (n = 12), ileo-colonic (n = 14) or colonic (n = 2) Crohn's disease were studied. In 25 out of 28 cases (89%) indication for surgery was strictures inducing an upper small bowel distension in 9 out of 25 patients. Mesenteric lymph nodes and liver biopsies, portal blood samples and peritoneum swabs were harvested after laparotomy and before gut opening. Bacterial translocation, defined as the presence of intestinal bacteria in at least one of the specimens, was present in 8 out of 28 patients. This was found in lymph nodes draining surgical territories in 7 out of 8 cases. Bacterial strains involved in translocation included E. coli (n = 5), Enterococcus (n = 3), Clostridium perfringens (n = 2), Proteus (n = 2), and Bacteroides fragilis (n = 1). The rate of translocation differed neither according to Crohn's disease site nor with perforating or non perforating type of the disease. Five out of 9 patients operated on for strictures with proximal distension had a translocation. In conclusion, bacterial translocation was identified in 29% of patients operated on for Crohn's disease in this series. Distension of the intestine proximal to a digestive stricture could favor the occurrence of bacterial translocation in Crohn's disease.

Adolescent

Bacterial translocation from the intestines.

Bacterial translocation is defined as the passage of viable bacteria from the gastrointestinal (GI) tract through the mucosal epithelium to other sites, such as the mesenteric lymph nodes, spleen, liver and blood. This paper reviews results from animal models utilized to obtain information concerning the defense mechanisms operating in the healthy host to confine bacteria to the GI tract. Gnotobiotic and antibiotic-decontaminated mice colonized with particular bacteria demonstrated that the indigenous GI flora maintains an ecologic equilibrium to prevent intestinal bacterial overgrowth and translocation from the GI tract. Studies with athymic (nu/nu) mice, thymus-grafted (nu/nu) mice, neonatally thymectomized mice, and mice injected with immunosuppressive agents demonstrated that the host immune system is another defense mechanism inhibiting bacterial translocation from the GI tract. Ricinoleic acid given orally to mice disrupted the intestinal epithelial barrier allowing indigenous bacteria to translocate from the GI tract. Thus, bacterial translocation from the GI tract of healthy adult mice is inhibited by: (a) an intact intestinal epithelial barrier, (b) the host immune defense system, and (c) an indigenous GI flora maintaining ecological equilibrium to prevent bacterial overgrowth. Deficiencies in host defense mechanisms act synergistically to promote bacterial translocation from the GI tract as demonstrated by animal models with multiple alterations in host defenses. Bacterial translocation occurred to a greater degree in mice with streptozotocin-induced diabetes, mice receiving nonlethal thermal injury, and mice receiving the combination of an immunosuppressive agent plus an oral antibiotic than in mice with only a primary alteration in host defenses. The study of bacterial translocation in these complex models suggests that opportunistic infections from the GI tract occur in discrete stages. In the healthy adult animal, bacterial translocation from the GI tract either does not occur or occurs at a very low level and the host immune defenses eliminate the translocating bacteria. Bacterial translocation does take place if one of the host defense mechanisms is compromised, such as a deficiency in the immune response, bacterial overgrowth in the intestines, or an increase in the permeability of the intestinal barrier. In this first stage, the bacteria usually translocate in low numbers to the mesenteric lymph node, and sometimes spleen or liver, but do not multiply and spread systemically.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Total parenteral nutrition promotes bacterial translocation from the gut.

Bacterial translocation from the gut may be the primary event in many disease processes. The purpose of this study was to examine the route of nutrient administration on bacterial translocation from the gut. Each of 90 female Fischer rats underwent placement of a central venous catheter and was randomized to one of three groups. Group I (control) received food and water ad libitum. Group II received standard TPN solution orally from a bottle sipper and drank the solution ad libitum. Group III underwent TPN via the central catheter by pair feeding of the animals with group II. Animals were fed for 2 weeks, and liver, spleen, mesenteric lymph nodes, blood, and cecum were aseptically obtained for culture. A statistically significant difference (p less than 0.014) was found between translocation rates of parenterally fed animals compared with enterally fed animals. Two thirds of the animals (18/27) fed parenterally had culture-positive mesenteric lymph nodes compared with one third (9/27) of the enterally fed group and none (0/30) of the control group. A statistically significant increase in the cecal bacterial count was demonstrated in the animals fed the TPN solution, independent of route. Parenteral nutrition promotes bacterial translocation from the gut by increasing the cecal bacterial count and impairing intestinal defense.

Animals

Does intravenous glutamine prevent bacterial translocation in hemorrhagic shock?

Bacterial translocation across the gut wall may be associated with insult to the latter. In this situation, intestinal flora can enter the blood stream and lymph nodes and be transported to other organs. Glutamine is a nonessential amino acid not presently included in total parenteral nutrition (TPN) preparations. The use of glutamine-enriched TPN in the rat has resulted in a significant reduction in bacterial translocation. This study attempted to evaluate the role of glutamine in preventing bacterial translocation following hemorrhagic shock in a rat model. Forty Sprague-Dawley rats were equally divided into two groups. The controls were given TPN solution, while the treated group had glutamine instead of the standard alanine present in TPN. Hemorrhagic shock was induced in both groups and blood cultures were performed. Glutamine-treated rats did not show a significant difference in survival suggesting that it is of no particular value in severe hemorrhagic shock in rats.

Alanine

The effect of intestinal transit time on bacterial translocation.

Increase in intraluminal bacterial count, disruption of the mucosal integrity, changes in intestinal immunity and transit time are the factors involved in bacterial translocation. The relationship between intestinal transit time, intra luminal bacterial count and translocation rate were investigated in 40 Wistar-albino rats. The study was conducted in 4 groups with 10 animals in each. Group I (controls): saline + laboratory chow, Group II: saline + oral total parenteral nutrition (TPN) solution, Group III: morphine sulfate (MS) + oral TPN solution, Group IV: neostigmine bromide (NB) + oral TPN solution. Intestinal transit time was measured by using Indium111-labeled diethylene triamine pentaacetic acid (DTPA). It was prolonged in the MS-treated group and shortened in the NB-treated group (p < 0.01). The frequency of bacterial translocation was 60% in the oral TPN solution group, 100% in the MS-treated group, 20% in the NB-treated group and 10% in controls. Bacterial counts in duodenum, jejunum, ileum and caecum were significantly increased (p < 0.001) in the MS-treated group and decreased (p < 0.05) in the NB-treated group in comparison with the control group. In conclusion, the prolongation of intestinal transit time increased the intraluminal bacterial count and augmented bacterial translocation. The decrease in intestinal transit time had a converse effect.

Animals

Bacterial translocation in colorectal cancers.

Bacterial translocation, the passage of viable indigenous bacteria from the gastrointestinal tract to the mesenteric lymph nodes and other internal organs, has been poorly studied in man to date. Pericolonic lymph nodes, liver, portal blood, and peritoneum specimens were harvested before antibiotics were administered during 20 operations for colorectal cancer and compared with those obtained in 20 operations for non colorectal conditions. Bacterial translocation, defined as the presence of intestinal bacteria in at least one of the specimens, was found in 13 patients (65 percent) in the colorectal cancer group as compared to 6 (30 percent) in the control group (p less than 0.05). The increased incidence of bacterial translocation in colorectal cancers was mainly due to the presence of bacteria in the pericolonic lymph nodes adjacent to the cancer. These findings suggest that intestinal bacteria translocate from the bowel lumen in a high proportion of patients with colorectal cancer and further stress the need for prophylactic antibiotics in colorectal cancer surgery.

Adenocarcinoma

[Bacterial translocation in multiple organ failure].

Bacterial translocation has been defined as the passage of both viable and nonviable bacteria and their products (e.g., endotoxins) across the intestinal barrier to extraintestinal sites such as the mesenteric lymph nodes, liver, etc. It has been hypothesized that intestinally derived bacteria or endotoxins serve as triggers to initiate, perpetuate, or exacerbate the septic state and thereby promote the development of multiple organ failure (MOF). In various animal studies, bacterial translocation has been associated with mortality and septic complications. Although most data on translocation have been derived from animal studies, convincing evidence has been provided that translocation may occur in humans during various disease states. The question still remains, however, of whether bacterial translocation is an important pathophysiological event in human disease or simply an epiphenomenon of severe disease, since the results are variable. Recent studies have indicated that the gut can produce important amounts of immunoinflammatory factors and that intestinal injury predisposes to distant organ injury even in the absence of detectable bacteria or endotoxins in the portal blood or tissues. This hypothesis may in part explain the inconsistent causal relationship between bacterial translocation and MOF.

Animals

The effects of G-CSF treatment and starvation on bacterial translocation in hemorrhagic shock.

BACKGROUND: Bacterial translocation is thought to be responsible for infectious complications after hemorrhagic shock. The aim of this study is to investigate the effects of granulocyte colony-stimulating factor (G-CSF) treatment on bacterial translocation in starved or fed animals subjected to hemorrhagic shock. MATERIALS AND METHODS: Fifty Wistar albino rats (200-275 g) were divided into six groups such as naive control (n = 7), G-CSF treatment (n = 7), hemorrhagic shock in starved rats (n = 9), hemorrhagic shock in fed rats (n = 9), G-CSF treatment 24 h before hemorrhagic shock in starved rats (n = 9), and G-CSF treatment 20 min after hemorrhagic shock in fed rats (n = 9). Hemorrhagic shock was induced by withdrawal of 2.1 ml/100 g blood via a carotid arterial cannulae placed under sodium pentobarbital anesthesia. Twenty-four hours later, mesenteric lymph nodes, liver, spleen, and peripheral blood samples were evaluated by using a quantitative microbiological technique and the numbers of colony-forming units were compared between groups. RESULTS: No bacteria was detected in samples from naive controls or G-CSF-treated unshocked rats. In animals subjected to hemorrhage, Escherichia coli was the predominant pathogen together with Streptococcus faecalis, Pseudomonas, and Lactobacillus species. In this model, starvation augmented the magnitude of bacterial translocation while G-CSF treatment has virtually abolished it. CONCLUSION: Under experimental conditions, preshock starvation increases gut-derived bacterial translocation and administration of G-CSF before or after hemorrhagic insult significantly reduces it.

Animals

Insulinlike growth factor 1 (IGF-1) reduces gut atrophy and bacterial translocation after severe burn injury.

Bacterial translocation after severe burns is associated with gut mucosal atrophy and increased mucosal permeability. Insulinlike growth factor 1 (IGF-1) levels are low after trauma and do not respond to growth hormone treatment. Since IGF-1 receptors have been demonstrated in gut mucosa, we proposed that treatment with IGF-1 would reduce mucosal atrophy and bacterial translocation. Rats received 50% total body surface area full-thickness burn or sham burn. They were treated with a continuous, subcutaneous infusion of either IGF-1 (approximately 3 mg/kg per day) or placebo (0.01 mol of acetate) for up to 5 days after receiving the burn. The mesenteric lymph node and liver were cultured for gram-negative bacteria. The small intestinal mucosa was scraped, weighed, and analyzed for DNA and protein content. Treatment with IGF-1 improved body weight, spleen weight, and gut mucosal weight. It stimulated mucosal DNA and protein content and reduced the incidence of bacterial translocation to the mesenteric lymph node from 89% to 30%. Insulinlike growth factor may reduce gut barrier failure by decreasing mucosal atrophy and subsequent barrier failure. In addition to its general anabolic effects, recombinant human IGF-1 may improve gut mucosal function and reduce infectious morbidity in severely traumatized or septic patients by reducing gut atrophy and reducing bacterial translocation.

Animals