Yersinia pseudotuberculosis infection with severe localised inflammation and ulceration of the ileum in a heart transplant patient.

In this case, we present an uncommon gastrointestinal infection in an immunocompromised patient that was solely diagnosed because of close collaboration between treating physicians and microbiologists. The patient is a 42-year-old male who underwent heart transplantation 5 years earlier. He presented with fever, weight loss, diarrhoea and tiredness. Initial investigations could not elucidate the aetiology of his symptoms. The patient was referred to the department of infectious diseases for further evaluation. Serology for Yersinia species was ordered and the result was suggestive for the possibility of a Yersinia species infection. Close collaboration between treating physicians and microbiologists followed and led to additional investigations, which revealed the diagnosis of a Yersinia pseudotuberculosis infection with extensive lesions in the gastrointestinal tract. Treatment with ciprofloxacin resulted in complete resolution of symptoms and healing of the gastrointestinal lesions. In conclusion, this case underlines the need for a multidisciplinary approach to complex patients of which symptoms have yet to be understood.

Bacterial translocation and mortality on rat model of intestinal ischemia and obstruction.

PURPOSE:: To develop an experimental model of intestinal ischemia and obstruction followed by surgical resection of the damaged segment and reestablishment of intestinal transit, looking at bacterial translocation and survival. METHODS:: After anesthesia, Wistar rats was subject to laparotomy, intestinal ischemia and obstruction through an ileal ligature 1.5cm of ileum cecal valve; and the mesenteric vessels that irrigate upstream of the obstruction site to approximately 7 to 10 cm were ligated. Abdominal wall was closed. Three, six or twenty-four hours after, rats were subject to enterectomy followed by an end to end anastomosis. After 24h, mesenteric lymph nodes, liver, spleen and lung tissues were surgically removed. It was studied survival rate and bacterial translocation. GraphPadPrism statistical program was used. RESULTS:: Animals with intestinal ischemia and obstruction for 3 hours survived 24 hours after enterectomy; 6hx24h: survival was 70% at 24 hours; 24hx24h: survival was 70% and 40%, before and after enterectomy, respectively. Culture of tissues showed positivity on the 6hx24h and negativity on the 3hx24h. CONCLUSION:: The model that best approached the clinic was the one of 6x24h of ischemia and intestinal obstruction, in which it was observed bacterial translocation and low mortality rate.

Bacterial translocation and mortality on rat model of intestinal ischemia and obstruction

Abstract Purpose: To develop an experimental model of intestinal ischemia and obstruction followed by surgical resection of the damaged segment and reestablishment of intestinal transit, looking at bacterial translocation and survival. Methods: After anesthesia, Wistar rats was subject to laparotomy, intestinal ischemia and obstruction through an ileal ligature 1.5cm of ileum cecal valve; and the mesenteric vessels that irrigate upstream of the obstruction site to approximately 7 to 10 cm were ligated. Abdominal wall was closed. Three, six or twenty-four hours after, rats were subject to enterectomy followed by an end to end anastomosis. After 24h, mesenteric lymph nodes, liver, spleen and lung tissues were surgically removed. It was studied survival rate and bacterial translocation. GraphPadPrism statistical program was used. Results: Animals with intestinal ischemia and obstruction for 3 hours survived 24 hours after enterectomy; 6hx24h: survival was 70% at 24 hours; 24hx24h: survival was 70% and 40%, before and after enterectomy, respectively. Culture of tissues showed positivity on the 6hx24h and negativity on the 3hx24h. Conclusion: The model that best approached the clinic was the one of 6x24h of ischemia and intestinal obstruction, in which it was observed bacterial translocation and low mortality rate.

Transcriptional Profiling of Ileocecal Valve of Holstein Dairy Cows Infected with Mycobacterium avium subsp. Paratuberculosis.

Johne's disease is a chronic infection of the small intestine caused by Mycobacterium avium subspecies paratuberculosis (MAP), an intracellular bacterium. The events of pathogen survival within the host cell(s), chronic inflammation and the progression from asymptomatic subclinical stage to an advanced clinical stage of infection, are poorly understood. This study examines gene expression in the ileocecal valve (ICV) of Holstein dairy cows at different stages of MAP infection. The ICV is known to be a primary site of MAP colonization and provides an ideal location to identify genes that are relevant to the progression of this disease. RNA was prepared from ICV tissues and RNA-Seq was used to compare gene transcription between clinical, subclinical, and uninfected control animals. Interpretation of the gene expression data was performed using pathway analysis and gene ontology categories containing multiple differentially expressed genes. Results demonstrated that many of the pathways that had strong differential gene expression between uninfected control and clinical cows were related to the immune system, such as the T- and B-cell receptor signaling, apoptosis, NOD-like receptor signaling, and leukocyte transendothelial migration pathways. In contrast, the comparison of gene transcription between control and subclinical cows identified pathways that were primarily involved in metabolism. The results from the comparison between clinical and subclinical animals indicate recruitment of neutrophils, up regulation of lysosomal peptidases, increase in immune cell transendothelial migration, and modifications of the extracelluar matrix. This study provides important insight into how cattle respond to a natural MAP infection at the gene transcription level within a key target tissue for infection.

Rapid testing and quantification of Salmonella in ileocaecal lymph nodes of Austrian pigs slaughtered for consumption.

Traditionally, quantitative microbial risk assessment (QMRA) is based on culture-dependent technologies. However, molecular quantification could forge additional, detailed information. A prerequisite of quantitative real-time PCR in animal science is a tissue preparation method where large volumes of tissue material can be reduced and particularly target cells can be concentrated. An easy-to-use sample preparation method for food (Matrix-Lysis) was recently adapted to tissues and now permits quantification of target cells from up to 5 g of organic matrix. The aim of this study was to examine the suitability of Matrix-Lysis for quantification of Salmonella in porcine ileocaecal lymph nodes (ICLNs). After demonstrating constant recovery rates, ICLNs from 540 pigs were examined for Salmonella spp. with Matrix-Lysis. Samples were also analysed using ISO 6579:2002, a combined enrichment/qPCR method and a lateral flow test. It could be shown that qPCR coupled with Matrix-Lysis can contribute to QMRA in food safety by enabling reproducible quantitative data, even at low contamination rates.

Análise da diversidade da microbiota intestinal de ratos submetidos à ressecção da valva ileocecal e criação de esfíncter artificial / Analysis of the diversity of the intestinal microbiota of rats subjected to resection of the ileocecalvalve and creation of artificial sphincter

OBJETIVO: analisar através de biologia molecular a diversidade da microbiota da junção ileocecal antes e após a ressecção da válvula ileocecal e reconstrução do trânsito com e sem a criação de "neoesfíncter". MÉTODOS: Os animais foram distribuídos em dois grupos: Grupo A (n=7) com ressecção da válvula ileocecal e anastomose ileocólica término-terminal em plano único, e Grupo B (n=7) com ressecção da válvula ileocecal e anastomose ileocólica término-terminal em plano único e confecção do esfíncter artificial. Reoperados com 20 dias coletou-se novamente conteúdo intraluminar do íleo e do cólon. Das amostras coletadas, extraiu-se DNA para reação de PCR-DGGE. Os padrões de bandas eletroforéticas , gerados na reação, foram submetidos ao programa Bionumerics para análise da similaridade e da diversidade da microbiota. RESULTADOS: a diversidade da microbiota foi maior e em mais amostras do íleo do que as do cólon. O grupo com a válvula apresentou os maiores valores e variações no cólon de 2,11 a 2,93. Em três animais de cada grupo estabeleceu-se comparação da similaridade e não se assemelharam ao controle. CONCLUSÃO: a ressecção da válvula ileocecal levou à mudanças da microbiota ileal e, com a criação de novo esfíncter, as variações foram maiores.

OBJECTIVE: To analyze, through molecular biology, the diversity of the intestinal microbiota before and after resection of the ileocecal junction and reconstruction of intestinal transit with and without the creation of a neosphincter. METHODS: Fourteen Wistar rats were divided into two groups: Group A (n = 7), submitted to resection of the ileocecal valve and end-to-end, single-layer ileocolic anastomosis; and Group B (n = 7) with resection of the ileocecal valve and end-to-end, single-layer ileocolic anastomosis followed by construction of an artificial sphincter. Intraluminal contents were collected from both groups. The animals were reoperated 20 days after the first procedure, with new collection of intraluminal contents of the ileum and colon. From the samples collected, DNA was extracted for PCR-DGGE. The electrophoretic banding patterns generated in the reaction were analyzed for similarities and diversities of the microbiota. RESULTS: The diversity of microorganisms was larger and in more samples when collected from the ileum than from the colon. The group with the neosphincter showed the highest variation in the colon, from 2.11 to 2.93. In three animals from each group was established comparing the similarity and not resembled the control. CONCLUSION: ileocecal resection led to changes in ileal microbiota and, with the creation of new sphincter, the changes were even greater.

Analysis of the diversity of the intestinal microbiota of rats subjected to resection of the ileocecal valve and creation of artificial sphincter.

OBJECTIVE: To analyze, through molecular biology, the diversity of the intestinal microbiota before and after resection of the ileocecal junction and reconstruction of intestinal transit with and without the creation of a neosphincter. METHODS: Fourteen Wistar rats were divided into two groups: Group A (n = 7), submitted to resection of the ileocecal valve and end-to-end, single-layer ileocolic anastomosis; and Group B (n = 7) with resection of the ileocecal valve and end-to-end, single-layer ileocolic anastomosis followed by construction of an artificial sphincter. Intraluminal contents were collected from both groups. The animals were reoperated 20 days after the first procedure, with new collection of intraluminal contents of the ileum and colon. From the samples collected, DNA was extracted for PCR-DGGE. The electrophoretic banding patterns generated in the reaction were analyzed for similarities and diversities of the microbiota. RESULTS: The diversity of microorganisms was larger and in more samples when collected from the ileum than from the colon. The group with the neosphincter showed the highest variation in the colon, from 2.11 to 2.93. In three animals from each group was established comparing the similarity and not resembled the control. CONCLUSION: ileocecal resection led to changes in ileal microbiota and, with the creation of new sphincter, the changes were even greater.

The small bowel flora in individuals with cecoileal reflux.

BACKGROUND: The observation of cecoileal reflux to barium enema is not rare; however, its causes and consequences have not been widely investigated. Considering that ileocecal junction exerts a function as barrier to invasion of bacteria from colon to small bowel, it seems interesting to study the intestinal microflora in subjects carrying cecoileal reflux. AIMS: This study aims at evaluating the ileal flora in individuals with cecoileal reflux. METHODS: A group of 36 subjects comprising 30 females and 6 males with a mean age of 54 years was assessed. Twenty-five individuals with cecoileal reflux and 11 without cecoileal reflux were submitted to small intestine contamination evaluation through the breath test with lactulose-H2 and measurement of the orocecal transit time by means of alternate current biosusceptometry. Small intestine bacterial overgrowth was characterized by orocecal transit time-H2 shortening. RESULTS: Comparison of basal H2, orocecal transit time-H2 and orocecal transit time-alternate current biosusceptometry measurements did not statistically differ between the groups with and without cecoileal reflux. Orocecal transit time-H2 was significantly smaller than orocecal transit time-alternate current biosusceptometry, particularly in individuals with cecoileal reflux. A significant correlation between the two methods was observed only in relation to control, not existing in relation to cecoileal reflux group. CONCLUSIONS: Smaller orocecal transit time-H2 and the loss of correlation with orocecal transit time-alternate current biosusceptometry observed in the individuals with cecoileal reflux suggest a differentiated behavior for such group relative to control, which could be associated with small intestine bacterial overgrowth.

The small bowel flora in individuals with cecoileal reflux / Estudo da flora do delgado em doentes com refluxo cecoileal

BACKGROUND: The observation of cecoileal reflux to barium enema is not rare; however, its causes and consequences have not been widely investigated. Considering that ileocecal junction exerts a function as barrier to invasion of bacteria from colon to small bowel, it seems interesting to study the intestinal microflora in subjects carrying cecoileal reflux. AIMS: This study aims at evaluating the ileal flora in individuals with cecoileal reflux. METHODS: A group of 36 subjects comprising 30 females and 6 males with a mean age of 54 years was assessed. Twenty-five individuals with cecoileal reflux and 11 without cecoileal reflux were submitted to small intestine contamination evaluation through the breath test with lactulose-H2 and measurement of the orocecal transit time by means of alternate current biosusceptometry. Small intestine bacterial overgrowth was characterized by orocecal transit time-H2 shortening. RESULTS: Comparison of basal H2, orocecal transit time-H2 and orocecal transit time-alternate current biosusceptometry measurements did not statistically differ between the groups with and without cecoileal reflux. Orocecal transit time-H2 was significantly smaller than orocecal transit time-alternate current biosusceptometry, particularly in individuals with cecoileal reflux. A significant correlation between the two methods was observed only in relation to control, not existing in relation to cecoileal reflux group. CONCLUSIONS: Smaller orocecal transit time-H2 and the loss of correlation with orocecal transit time-alternate current biosusceptometry observed in the individuals with cecoileal reflux suggest a differentiated behavior for such group relative to control, which could be associated with small intestine bacterial overgrowth.

RACIONAL: Fato de observação não rara, é o encontro de refluxo cecoileal durante realização de enema opaco. As causas e conseqüências deste achado têm sido pouco estudadas. OBJETIVOS: Sabendo que a junção ileocecal exerce função de barreira e proteção contra a invasão do delgado pela flora colônica, realizou-se o presente estudo com a finalidade de investigar se existe contaminação ileal em indivíduos com refluxo cecoileal ao enema opaco. MÉTODOS: Investigaram-se 36 indivíduos, 30 mulheres e 6 homens, idade média de 54 anos, 25 com e 11 com ausência refluxo cecoileal. Todos submetidos a pesquisa de contaminação bacteriana do delgado por intermédio de teste respiratório com lactulose-H2 e a determinação do tempo de trânsito orocecal por meio de biossusceptometria de corrente alternada. A caracterização da contaminação do delgado foi baseada no encurtamento do tempo de trânsito orocecal medido pelo teste da lactulose-H2. RESULTADOS: A comparação dos valores basais do H2, do tempo de trânsito orocecal-H2 e tempo de trânsito orocecal-biossusceptometria de corrente alternada não diferiram estatisticamente entre os grupos com e sem refluxo cecoileal. Quando comparados os tempo de trânsito orocecal-H2 e tempo de trânsito orocecal-biossusceptometria, foi observado aumento de tendência de redução do primeiro em relação ao último nos grupos com refluxo cecoileal e correlação significante entre os dois métodos apenas no grupo-controle, inexistindo nos com refluxo cecoileal. CONCLUSÃO: Encurtamento do tempo de trânsito orocecal-H2 e sua perda de correlação com o tempo de trânsito orocecal-biossusceptometria observado em indivíduos com refluxo cecoileal, sugerem comportamento diferenciado deste grupo em relação ao grupo-controle. Possível explicação para as diferenças registradas entre os grupos, seria a presença de flora anômala nos indivíduos com refluxo cecoileal.

[Pseudotumoral form of abdominal tuberculosis: report of four cases]. / La tuberculose abdominale pseudo-tumorale.

PURPOSE: To report the imaging features of four cases of pseudo-tumoral abdominal tuberculosis in order to demonstrate diagnostic difficulties, even in endemic areas, despite expanding imaging modalities. MATERIAL AND METHODS: We have retrospectively reviewed four cases investigated in our hospital between 1998 and 2002. The first case concerned a 2 year and 6 month old boy admitted for a right iliac fossa mass. The other patients were 43 and 48 year old women admitted for epigastric mass and persistent epigastric pain with past history of treated tuberculosis respectively. The last was an 86 year old man investigated because progressive worsening of general health. Sonography (US) and computed tomography (CT) were performed in all four cases. RESULTS: Involvement of the ileocaecal region was noted in three cases. Large intestinal hypervascular mass in the pediatric case, heterogeneous mass with porta hepatis lymph node in the patient with past history of treated tuberculosis, pseudotumoral caecal wall thickening in the third case. The multiloculated pattern of the epigastric mass and its location initially suggested a diagnosis of pancreatic tumor. CONCLUSION: Abdominal tuberculosis is rare even in endemic areas. This diagnosis must be considered when imaging modalities depict a hypervascular soft tissue mass on Doppler examination with peripheral enhancement on CT scan.

[Incidence of bacterial translocation in four different models of experimental short bowel syndrome]. / Incidencia de la traslocación bacteriana en cuatro modelos diferentes de intestino corto experimental.

UNLABELLED: The outcome of patients with short bowel syndrome is influenced for factors such as the length of remnant intestine or the presence or absence of ileocecal valve (ICV). Gram-negative sepsis, the main cause of mortality in this group of children, is probably due to bacterial translocation (BT), because after gut resection there are a number of circumstances that favour its occurrence, being the most known intestinal dismotility, bacterial overgrowth, loss of gut-associated lymphoid tissue, total parenteral nutrition (TPN) and fasting related mucosal atrophy. The aim of this experimental controlled study was to test the incidence of BT after four different types of gut resection, in animals fed orally or receiving TPN. Hundred and three adult Wistar rats bred and raised in our facilities according to European Union Regulations were randomly divided in six groups:--Group 1 (n = 26): non-manipulated animals, served as a control.--Group 2 (n = 14): 80% non-lethal small bowel resection, fed orally.--Group 3 (n = 15): same resection as group 2 but including ICV. Rat chow ad libitum.--Group 4 (n = 27): non-resected fasting animals receiving all-in-one TPN solution.--Group 5 (n = 11): same resection as group 2, but fasting and receiving TPN--Group 6 (n = 10): 90% small bowel resection, including cecum and ICV, fasting and TPN. The animals were maintained for 10 days in individual metabolic cages, and, at the end of the experiment, were bled by portal and cardiac puncture. Mesenteric lymph nodes, peripheral and portal blood samples were cultured for BT. Non-manipulated rats (group 1) had lower BT incidence (8%) than resected ones (groups 2, 3, 5 and 6, 93%, 60%, 91%, 60%, p < 0.05) or animals non-resected, receiving TPN (group 4.51%, p < 0.05). When resection included ICV in orally fed rats BT index was also lower (group 3 vs group 2.60% vs 91%, p < 0.05). In TPN resected animals a drop was also found in BT when ICV and cecum were added to small bowel resection (group 6 vs group 5.60% vs 91%, p < 0.05). IN CONCLUSION: 1. Gut resection is associated to a high degree of BT, even if the animals are fed orally. 2. Resection including ICV, produced less BT. 3. TPN-related BT was shown in half of the animals non resected. 4. TPN-resected rats had also less BT when ICV and cecum were removed.

Progressive bovine paratuberculosis is associated with local loss of CD4(+) T cells, increased frequency of gamma delta T cells, and related changes in T-cell function.

Bovine paratuberculosis is caused by the infection of young calves with Mycobacterium avium subsp. paratuberculosis, resulting in a chronic granulomatous infection of predominantly the ileum. After an incubation period of 2 to 5 years, the disease becomes progressive in some of the chronically infected, but asymptomatic cows. This results in a protein-losing enteropathy that will ultimately be fatal. A loss of cell-mediated immune responses in symptomatic animals has been described, but no information is available concerning immune reactivity in the intestine. We sought to investigate putative disease status-associated lymphocyte subset distributions and antigen-specific functional characteristics of mononuclear cells isolated from blood, gut-associated lymphoid tissue, and the intestinal walls of 22 cows in different stages of disease and in control animals. The results demonstrated a significant decrease in CD4(+) T-cell frequency and a significant increase in TcR1-N12(+) gamma delta T-cell frequency in ileum lamina propria lymphocytes of symptomatic animals compared to the asymptomatic shedders. Immunohistology revealed that there was also an absolute decrease in the number of CD4(+) T cells in sections of the lesional ileum. Our findings also indicated that both peripheral and intestinal cell-mediated responses are decreased in symptomatic animals compared to asymptomatic animals. We conclude that the decrease in cell-mediated responses is likely related to a loss of antigen-specific CD4(+) T cells, which is most prominent in the lesional ileum from symptomatic animals, thus contributing to the progressive nature of bovine paratuberculosis.

[Bacterial translocation associated with short bowel: role of ileocecal valve and cecum]. / La traslocación bacteriana asociada al intestino corto: papel de la válvula ileocecal y del ciego.

Sepsis in short bowel syndrome (SBS) is due in part to bacterial translocation (BT). Parenteral nutrition (PN) is often necessary in SBS and promotes BT. The presence of ileocecal valve (ICV) has been considered as a good prognostic factor in the outcome of this children. The aim of this study was to asses the effect of the presence or absence of ICV and cecum in five different models of gut resection in the rat. Fifty-five adult Wistar rats were randomly assigned to one of five groups: Group 1 (N = 14): standard rat chow + 80% small bowel resection. Group 2 (N = 10): standard rat chow + 80% small bowel resection including cecum. Group 3 (N = 10): standard rat chow + 80% small bowel resection including ICV. Group 4 (N = 11): NP + 80% small bowel resection. Group 5 (N = 10): NP + 80% small bowel resection including ICV and cecum. Ten days after surgery they were sacrificed and mesenteric lymph nodes (MLN), spleen and peripheral (PBL) and portal blood (POBL) specimens were recovered and cultured. Groups 3 (without ICV, with cecum) and 5 (without ICV, without cecum) showed 60% BT in MLN and POBL, and groups 1 and 4 (with ICV, without cecum) 93% and 91% respectively (p < 0.05). In PBL, group 3 (without ICV, with cecum) showed also less BT than groups 1 and 4 (10% vs 43% and 55% respectively, p < 0.05) and group 5 (without ICV and cecum) had less BT than groups 1, 2 and 4 (0% vs 43%, 30% and 55%, p < 0.01). In conclusion, these results suggest that the absence of ICV decreases BT and that the cecum does not seems to play a role on his.

[Gastrointestinal tuberculosis--an almost forgotten clinical picture]. / Gastrointestinalis tuberculosis--egy majdnem elfeledett kórkép.

Incidence of gastrointestinal tuberculosis increases in the western world. Exact epidemiological data about this rare extrapulmonological disease are not available in Hungary. Three advanced cases with intestinal tuberculosis have been reported, all of them recognised during surgery due to complications. The clinicopathological correlations are briefly reviewed, and the importance of early diagnosis is emphasized.

[An intestinal neosphincter attained by circumscribed muscle proliferation. Technical development and functional evaluation in the dog]. / Ein intestinaler Neosphinkter (INS) durch umschriebene Muskelvermehrung. Technische Entwicklung und funktionelle Evaluierung beim Hund.

In an experimental study an intestinal neosphincter (INS) was constructed by modifying the principle of the ileocolic nipple-valve anastomosis by means of ultrasonic tissue fragmentation of the contacting serosa of the ileum and the corresponding mucosa of the ileum and colon. The healing of the muscle layers was studied histologically. The function of the INS was investigated in six dogs and compared intraindividually with that of the ileocecal valve and conventional end-to-end anastomosis. Morphologically the neospincters healed within 3 months without major fibrosis. The reference values of the aerobic and anaerobic bacterial counts in the terminal ileum were more than 2 logs lower than in the colon with the normal ileocecal valve, and after ileo-colonic end-to-end-anastomosis bacterial colonization of the terminal ileum was found both qualitatively and quantitatively. Subsequent interposition of the INS led to bacterial clearance of the terminal ileum. The median aerobic bacterial counts were lower by six logs and the an aerobic bacterial counts by 3 logs than in the colon. However, differences were not statistically significant owing to the wide variation in the individual values. Nevertheless, the demonstrable clearance of the terminal ileum could be explained by the orthograde passage with absolutely no stagnation and the relative competence of the INS in resisting retrograde pressure competence. In conclusion, ultrasonic fragmentation of the serosa and mucosa of the bowel allows construction of an INS from three muscle layers, which acts as a bacteriological barrier. Before it is introduced into the clinical setting its integration into the intestinal motility should be evaluated by further studies.