Elevated Fecal Candida Counts in Patients with Antibiotic-Associated Diarrhea: Role of Soluble Fecal Substances

To assess the role of soluble fecal substances in the elevation of fecal Candida counts in patients with antibiotic-associated diarrhea (AAD), we investigated the growth of Candida albicans in vitro in serially diluted stool fluids from patients with AAD and healthy subjects. There were significantly higher Candida albicans counts in stool fluids diluted 1:10 from AAD patients than in healthy subjects and the phosphate-buffered saline growth control, which may be due to reduced soluble Candida inhibitors and increased availability of growth factors and nutrients.     

Candida and antibiotic-associated diarrhoea

The role of Candida in antibiotic-associated diarrhoea (AAD) has been controversial for many years. Since Candida exists physiologically in the gastrointestinal tract, the presence of small numbers of Candida organisms in stool has therefore been considered normal, and thus non-pathogenic. Increased Candida counts have been linked to the development of diarrhoea in antibiotic-treated patients. However, recent findings have not confirmed this. To date, there is no convincing evidence that Candida may cause AAD in adults.     

Differences in fecal microflora between patients with atopic dermatitis and healthy control subjects

BACKGROUND: The prevalence of allergic diseases, such as atopic dermatitis (AD), has been increasing. However, few investigations have been made of the intestinal microflora in Japanese patients with AD. OBJECTIVE: The purpose of this study was to determine the differences in microflora, fecal serum IgA concentrations, and skin IgA contents between patients with AD and healthy control subjects. METHODS: This trial was conducted as a case-control study using 30 minor patients with AD and age- and sex-matched healthy control subjects (n = 68). One week after a questionnaire was administered, fecal specimens and 24-hour skin secretion specimens were collected from all subjects. Fecal microflora, fecal IgA concentrations, and IgA contents on the skin surface were analyzed. RESULTS: The counts of Bifidobacterium (in log10 colony-forming units per gram) were significantly lower in patients with AD than in healthy control subjects (9.75 +/- 0.68 vs 10.10 +/- 0.50 log(10) colony-forming units/g, P <.05). In particular, percentages of Bifidobacterium were significantly lower in patients with severe skin symptoms than in those with mild skin symptoms (40% +/- 6% vs 19% +/- 6%, P <.05). In addition, the frequency of occurrence of Staphylococcus was significantly higher in patients with AD than in healthy control subjects (83% vs 59%, P <.05). There were no significant differences in fecal IgA content or IgA content on the skin between the 2 groups. CONCLUSION: Patients with AD had lower counts of Bifidobacterium than healthy control subjects, and the frequency of Staphylococcus was higher in patients with AD than in control subjects. Disorder of the intestinal microflora might play a role in the onset of AD and the aggravation of skin symptoms.     

Enrichment of Multilocus Sequence Typing Clade 1 with Oral Candida albicans Isolates in Patients with Untreated Periodontitis

This study investigated the prevalence and cell density of Candida species in periodontal pockets, healthy subgingival sites, and oral rinse samples of patients with untreated periodontitis. Twenty-one periodontitis patients underwent sampling at two periodontitis sites, and 19/21 of these patients underwent sampling at one periodontally healthy site. Both paper point and curette sampling techniques were employed. The periodontitis patients and 50 healthy subjects were also sampled by oral rinse. Candida isolates were recovered on CHROMagar Candida medium, and representative isolates were identified. Candida spp. were recovered from 10/21 (46.7%) periodontitis patients and from 16/50 (32%) healthy subjects. C. albicans predominated in both groups and was recovered from all Candida-positive subjects. Candida-positive periodontitis patients yielded Candida from periodontal pockets with average densities of 3,528 and 3,910 CFU/sample from curette and paper point samples, respectively, and 1,536 CFU/ml from oral rinse samples. The majority (18/19) of the healthy sites sampled from periodontitis patients were Candida negative. The 16 Candida-positive healthy subjects yielded an average of 279 CFU/ml from oral rinse samples. C. albicans isolates were investigated by multilocus sequence typing (MLST) to determine if specific clonal groups were associated with periodontitis. MLST analysis of 31 C. albicans isolates from periodontitis patients yielded 19 sequence types (STs), 13 of which were novel. Eleven STs belonged to MLST clade 1. In contrast, 16 C. albicans isolates from separate healthy subjects belonged to 16 STs, with 4 isolates belonging to clade 1. The distributions of STs between both groups were significantly different (P = 0.04) and indicated an enrichment of C. albicans isolates in periodontal pockets, which warrants a larger study.     

The prevalence of Candida albicans-associated diarrhoea in Buea, South West Cameroon

A total of 362 stool specimens were collected from 184 and 178 patients presenting at the Buea district Hospital with and without diarrhoea, respectively. The samples were screened and cultured for Candida albicans using standard microbiological procedures. Of the 184 diarrhoeic stool cultures, 35.9% showed C. albicans overgrowth as indicated by count >or=10(4) CFL/mL. Of the 178 non diarrhoeic stool cultures, C. albicans was identified in 23.6% of samples and counts were all <10(4) CFU/mL. An association was observed for C. albicans overgrowth and diarrhoea (p<0.001). The majority of isolates (87.8%) from the 66 samples showing candida overgrowth were susceptible to Amphotericin B in anti-fungal drug sensitivity assays. Results of the study highly suggest that C. albicans is an important cause of diarrhoea in the study area. We recommend that this fungus should be routinely checked in individuals presenting with diarrhoea particularly children and patients on prolonged or frequent antibiotic therapy.     

The magnetic immuno polymerase chain reaction assay for direct detection of salmonellae in fecal samples.

Direct polymerase chain reaction (PCR)-based detection with fecal specimens is hampered by inhibitory compounds, such as bilirubin and bile salts. These fecal compounds showed significant inhibition of PCR at low concentrations (10 to 50 micrograms/ml). For direct PCR analysis, fecal samples must be diluted 500-fold to overcome inhibition. Therefore, the magnetic immuno PCR assay (MIPA), which combines immunomagnetic separation by using specific monoclonal antibodies and PCR, was used to directly detect salmonellae in feces from humans. Immunomagnetically extracted stool samples needed to be diluted only 10-fold when 1 microgram of T4 gene 32 protein was added to the PCR. The MIPA sensitivity obtained was 10(5) CFU/ml of feces. A panel of monoclonal antibodies specific for Salmonella serogroups A to E was used to extract salmonellae from clinical samples. MIPA detection of salmonellae occurred with 11 out of 14 stool samples stored at 4 degrees C for 2 months. MIPA detection of salmonellae in stool samples is a promising, fast method for detection and identification.     

Multilocus sequence typing reveals intrafamilial transmission and microevolutions of Candida albicans isolates from the human digestive tract

Candida albicans is a human commensal that is also responsible for superficial and systemic infections. Little is known about the carriage of C. albicans in the digestive tract and the genome dynamics that occur during commensalisms of this diploid species. The aim of this study was to evaluate the prevalence, diversity, and genetic relationships among C. albicans isolates recovered during natural colonization of the digestive tract of humans, with emphasis on Crohn's disease patients who produce anti-yeast antibodies and may have altered Candida sp. carriage. Candida sp. isolates were recovered from 234 subjects within 25 families with multiple cases of Crohn's disease and 10 control families, sampled at the oral and fecal sites. Prevalences of Candida sp. and C. albicans carriage were 53.4% and 46.5%, respectively, indicating frequent commensal carriage. No differences in prevalence of carriage could be observed between Crohn's disease patients and healthy subjects. Multilocus sequence typing (MLST) of C. albicans isolates revealed frequent colonization of a subject or several members of the same family by genetically indistinguishable or genetically close isolates. These latter isolates differed by loss-of-heterozygosity events at one or several of the MLST loci. These loss-of-heterozygosity events could be due to either chromosome loss followed by duplication or large mitotic recombination events between complementary chromosomes. This study was the first to jointly assess commensal carriage of C. albicans, intrafamilial transmission, and microevolution. The high frequency of each of these events suggests that the digestive tract provides an important and natural niche for microevolutions of diploid C. albicans through the loss of heterozygosity.     

Measurement of fecal lactoferrin as a marker of fecal leukocytes.

While diarrheal illnesses are extremely common in communities and hospitals throughout the world, an etiologic diagnosis may be expensive and cost-ineffective. Although the presence of fecal leukocytes are helpful in the diagnosis and specific therapy of inflammatory diarrheas, this requires prompt microscopic examination of fecal specimens (preferably obtained in a cup rather than a swab or diaper) by a trained observer. We developed a simple, sensitive test for the detection of leukocytes in fecal specimens using antilactoferrin antibody. Whereas radial immunodiffusion detected 0.02 micrograms of lactoferrin (LF) per microliter or greater than or equal to 2,000 leukocytes per microliter, latex agglutination (LA) readily detected greater than or equal to 0.001 micrograms of LF per microliter or greater than or equal to 200 leukocytes per microliter added to stool specimens. Despite the destruction or loss of morphologic leukocytes on storage for 1 to 7 days at 4 degrees C or placement of specimens on swabs, measurable LF remained stable. Initial studies of stool specimens from six patients with Salmonella or Clostridium difficile enteritis were positive and those from three controls were negative for LF by LA. Of 17 children in Brazil with inflammatory diarrhea (greater than or equal to 1 leukocyte per high-power field), 16 (94%) had LF titers of greater than 1:50 by LA, whereas only 3 of 12 fecal specimens with less than 1 leukocyte per high-power field on methylene blue examination and none of 7 normal control specimens had an LF titer of greater than 1:50 by LA. Of 16 fecal specimens from patients with C. difficile diarrhea (cytotoxin titers, >/= 1:1,000), 95% (n = 15) had detectable LF by LA (in titers of 1:100 to 1: 800). Finally, of 48 fecal specimens from healthy adult U.S. volunteers before and after experimental shigellosis and of 29 fecal specimens from children with documented shigellosis and hospitalized controls in northeastern Brazil, fecal LF titers ranged from 1:200 to >/= 1:5,000 in 96% (25 of 26) samples from patients with shigellosis (and reported positive for fecal leukocytes), while 51 controls consistently had fecal LF titers of 相似文献   

Alteration of T cell function in healthy persons with a history of thymic x-irradiation.

The possible late effects of x-irradiation to the infantile thymus were investigated by studying immune functions in 12 healthy persons with a history of thymic x-irradiation and healthy control subjects. No differences were found in serum immunoglobulin values, humoral antibody levels, lymphocyte counts, and lymphocyte reactivity to phytochemagglutinin, vaccinia virus, purified protein derivative (PPD), and allogeneic cells. The irradiation group exhibited cellular hyperresponsiveness to streptoskinase-streptodornase (SK-SD). In contrast, mean skin and in vitro lymphocyte responses to Candida albicans were depressed in the patients with thymic irradiation. A dissociation of these two Candida responses was found in only 1 of 14 healthy control subjects but in 7 of 12 irradiated individuals. While thymic irradiation did not result in impaired immunologic defenses leading to clinical disease, it caused alterations in T cell responses similar to those reported in patients with chronic mucocutaneous candidiasis.     

Candida and Saccharomyces spp. fungal associations in fecal microbiocenosis of diabetes patients and healthy subjects

The article is devoted to analysis of pathogenic and diagnostic significance of Candida and Saccharomyces co-existence in diabetic patients. These transient fungi are known to be present in fecal microbiocenosis of both healthy subjects and patients with diabetes mellitus. However, their overall occurrence is significantly increased in the disease and the structure of the biocenosis undergoes alteration. These data confirm the role of yeast-like fungi in pathogenesis of diabetes. The diagnostic value of detection of monospecific and mixed populations of Candida and Saccharomyces spp. is not very high, but their presence in feces, especially in women, may be regarded as a sign of disturbed carbohydrate metabolism.     

Gastrointestinal tract cancer screening using fecal carcinoembryonic antigen

There is a great need to detect gastrointestinal tract cancer at an early stage. It is well known that most carcinoma tissues of the gastrointestinal tract contain carcinoembryonic antigen (CEA). Stools are a rich source of cells derived from the gastrointestinal tract. We analyzed total fecal CEA in 60 gastrointestinal tract cancer patients, 20 benign gastrointestinal tract disorder patients, and 240 normal controls, using a simple, reliable method. We compared the sensitivity and specificity of fecal CEA with those of serum CEA and fecal occult blood test (FOBT). The level of fecal CEA in gastrointestinal tract cancer was much higher than controls (44.1 +/- 70.1 ng/mg stool vs 3.7 +/- 3.5 ng/mg stool, p < 0.001) and was not increased in benign gastrointestinal disorders (4.5 +/- 8.2 ng/mg stool). Fecal CEA level was > 10 ng/mg stool in 22 of 32 samples (69%)from stomach cancer patients and 24 of 28 samples (86%)from colorectal cancer patients. The sensitivity of serum CEA (> 5 ng/ml) was 19% in stomach cancer and 39% in colorectal cancer, whereas the sensitivity of FOBT was 13% in stomach cancer and 21% in colorectal cancer. The specificity of fecal CEA was 90% in benign gastrointestinal tract disorders and 93% in normal controls. This specificity was similar to those of serum CEA and FOBT. In conclusion, fecal CEA measurement is superior to serum CEA or FOBT for detection of gastrointestinal tract cancer. Fecal CEA may become the screening test of choice for gastrointestinal tract cancer.     

Elevated Candida antigen titers are associated with neutrophil dysfunction after injury.

This study was undertaken to determine if impaired neutrophil (polymorphonuclear leukocytes [PMNL]) function is associated with an elevated Candida antigen titer after injury. PMNL from eight severely injured adults with Candida antigen titers of > or = 1:4 (titer positive) were evaluated for the ability to inhibit growth of Candida albicans in vitro by using a [3H]glucose incorporation assay. PMNL from eight severely injured adults with titers of < 1:4 (titer negative) and from eight healthy volunteers were studied for comparison. PMNL from the titer-positive patients had suppressed ability to inhibit C. albicans growth compared with PMNL from titer-negative patients and healthy volunteers. In vitro, PMNL function against C. albicans could be augmented significantly by cytokines. Granulocyte macrophage-colony-stimulating factor was most potent at augmenting function, followed by interleukin-8 and gamma interferon. Injured patients with elevated candida antigen titers have impaired PMNL function against C. albicans, and this function can be restored by cytokines.     

Rapid fecal cytokeratin-19 test and fecal occult blood test in screening for gastrointestinal diseases

To evaluate the screening power of the fecal cytokeratin-19 test (CK-19) and the fecal occult blood test (FOBT), we performed rapid fecal CK-19 and FOBT tests on 515 stool samples from patients with various GI diseases and 814 stool samples from control patients. The rapid fecal CK-19 test (developed by DiNonA Research Institute, Seoul, Korea) is based on gold immunochromatography and has a sensitivity of 1 ng/ml. The positive rate of the FOBT was 2.1% in controls, 14.0% in GI cancer patients, 3.5% in GI inflammation patients, 11.7% in bone marrow transplant (BMT) patients, and 6.0% in childhood diarrhea patients. Except for the GI inflammation patients, the patients' positive rates for FOBT were all higher than the controls (p <0.05). The positive rate of the fecal CK-19 test was 8.2% in controls, 42.1% in GI cancer patients, 66.0% in GI inflammation patients, 84.8% in BMT patients, and 19.9% in childhood diarrhea patients. In all of the patient groups, positive rates for the CK-19 test were higher than in the controls (p <0.05). The fecal CK-19 test was more frequently positive (42.1%) in GI cancer patients than the FOBT; if both tests were used, the sensitivity was 49.1%. The fecal CK-19 test (but not the FOBT) gave a higher positive rate in GI inflammation patients than the controls, suggesting that the CK-19 test could serve as a screening test for GI inflammation. The highest positive rate of the fecal CK-19 test was found in the BMT group, indicating that significant GI epithelial desquamation had occurred. Although the positive rate of the fecal CK-19 test in childhood diarrhea patients was higher than in the controls, it was much lower than in adults with GI inflammatory disease. Evidently, children with GI inflammation do not desquamate as much intestinal epithelium as adult patients with GI inflammation. This study shows that the rapid fecal GK-19 test, used in conjunction with the FOBT, may be a valuable screening technique for GI diseases and can assist physicians in the differential diagnosis of GI diseases.     

Use of fermenter-dialysis culture for the cultivation of medically relevant microbes. III. Candida albicans

For mass cultivation of Candida albicans (serotype A), a fermenter dialysis culture technique is described and compared with shaking culture and fermenter batch culture techniques. Important growth parameters such as yeast dry weight and viable cell counts demonstrate the advantage of the fermenter dialysis culture. Mannan, the major antigen from Candida albicans prepared by phenol-water extraction followed by gel chromatography was tested with the monoclonal IgM antibody H5.     

Anticandidal activity and interleukin-1 beta and interleukin-6 production by polymorphonuclear leukocytes are preserved in subjects with AIDS.

Polymorphonuclear granulocytes (PMN; or neutrophils) from uninfected or human immunodeficiency virus-infected subjects were tested for their ability to inhibit growth of Candida albicans and produce interleukin-1 beta (IL-1 beta) and IL-6 in vitro. It was seen that PMN from AIDS (Centers for Disease Control stage IV) patients expressed equal if not greater anticandidal activity compared with the activity expressed by neutrophils from all other subjects examined. On exposure to granulocyte macrophage-colony-stimulating factor or to a mannoprotein constituent (MP-F2) from C. albicans itself, PMN from AIDS patients showed enhanced antifungal activity and production of remarkable quantities of IL-1 beta and IL-6. These findings suggest that the functional abilities of PMN to inhibit Candida growth and secrete relevant proinflammatory and immunomodulatory cytokines are intrinsically preserved in AIDS patients.     

Increased levels of Candida albicans mannan-specific T-cell-derived antigen binding molecules in patients with invasive candidiasis

In addition to cytokines, CD4+ T cells have been found to secrete soluble, T-cell-derived antigen binding molecules (TABMs). These antigen-specific immunoproteins are thought to have immunoregulatory properties in the suppression of cell-mediated immunity (CMI) because they often associate with interleukin-10 (IL-10) and transforming growth factor beta. Decreased CMI causes susceptibility to infections caused by organisms which are normally nonpathogenic. In this situation, e.g., Candida albicans saprophytism may develop into invasive candidiasis. The difficult diagnosis of invasive candidiasis is based on the findings obtained from blood cultures and with tissue biopsy specimens, with some additional diagnostic value gained by the detection of Candida albicans mannan antigenemia and antimannan antibodies. In the present study, Candida albicans mannan-specific TABM (CAM-TABM) levels in the sera of patients with invasive candidiasis (n = 11), Candida colonization (n = 11) and noncolonization (n = 10), recurrent vulvovaginal candidiasis (n = 30), and atopic eczema dermatitis syndrome (n = 59) and healthy controls (n = 30) were analyzed. For 14 participants, the effect of mannan stimulation on TABM production and gamma interferon (IFN-gamma) and IL-4 mRNA expression by peripheral blood lymphocytes was also studied. It was demonstrated that CAM-TABM production was the highest in patients with invasive candidiasis and that CAM-TABM levels could distinguish Candida-colonized patients from noncolonized patients. In addition, the CAM-TABM level was directly related to mRNA expression for IL-4 but not IFN-gamma. These results reinforce the view that TABMs are associated with decreased CMI, immunoregulation, and the T-helper cell 2-type immune response.     

Isolation of Vibrio parahaemolyticus from fecal specimens on mannitol salt agar.

Normal sera and sera from burned patients were examined for Candida agglutinin titers, precipitin titers, and the ability to disperse germ tubes of Candida albicans in an attempt to determine whether germ tube dispersion is correlated with Candida infection as animal models have indicated. Other investigators have reported that immunoglobulin G antibody to Candida interferes with a serum clumping factor resulting in germ tube dispersion. Germ tube dispersion in sera from burned patients with varying degrees of Candida infection is significantly greater than that found in uninfected controls. In addition, the germ tube dispersion test indicated the presence of Candida infection in several patients who had clinical evidence of infection but no detectable agglutinins or precipitins.     

One-time screening for colorectal cancer with combined fecal occult-blood testing and examination of the distal colon

BACKGROUND: Fecal occult-blood testing and sigmoidoscopy have been recommended for screening for colorectal cancer, but the sensitivity of such combined testing for detecting neoplasia is uncertain. At 13 Veterans Affairs medical centers, we performed colonoscopy to determine the prevalence of neoplasia and the sensitivity of one-time screening with a fecal occult-blood test plus sigmoidoscopy. METHODS: Asymptomatic subjects (age range, 50 to 75 years) provided stool specimens on cards from three consecutive days for fecal occult-blood testing, which were rehydrated for interpretation. They then underwent colonoscopy. Sigmoidoscopy was defined in this study as examination of the rectum and sigmoid colon during colonoscopy, and sensitivity was estimated by determining how many patients with advanced neoplasia had an adenoma in the rectum or sigmoid colon. Advanced colonic neoplasia was defined as an adenoma 10 mm or more in diameter, a villous adenoma, an adenoma with high-grade dysplasia, or invasive cancer. Classification of subjects according to the findings was based on the most advanced lesion. RESULTS: A total of 2885 subjects returned the three specimen cards for fecal occult-blood testing and underwent a complete colonoscopic examination. A total of 23.9 percent of subjects with advanced neoplasia had a positive test for fecal occult blood. As compared with subjects who had a negative test for fecal occult blood, the relative risk of advanced neoplasia in subjects who had a positive test was 3.47 (95 percent confidence interval, 2.76 to 4.35). Sigmoidoscopy identified 70.3 percent of all subjects with advanced neoplasia. Combined one-time screening with a fecal occult-blood test and sigmoidoscopy identified 75.8 percent of subjects with advanced neoplasia. CONCLUSIONS: One-time screening with both a fecal occult-blood test with rehydration and sigmoidoscopy fails to detect advanced colonic neoplasia in 24 percent of subjects with the condition.     

Suppression of fungal growth exhibited by Pseudomonas aeruginosa.

Three surgery patients were monitored postoperatively, with particular reference to lung infection. In each case there was a clinical impression that Pseudomonas aeruginosa suppressed the growth of Candida albicans in patients with clinically significant lung infections from whom both of these organisms were isolated from serial sputum samples. Regrowth of C. albicans after P. aeruginosa eradication occurred in two patients, despite fluconazole therapy, to which both C. albicans isolates were susceptible. In all three patients, the strain of P. aeruginosa was found to inhibit the growth of the corresponding C. albicans strain in vitro. Further in vitro susceptibility studies revealed significant inhibition by 10 strains of P. aeruginosa of 11 strains of fungi known to infect humans; these were Candida krusei, Candida keyfr, Candida guillermondii, Candida tropicalis, Candida lusitaniae, Candida parapsilosis, Candida pseudotropicalis, Candida albicans, Torulopsis glabrata, Saccharomyces cerevisiae, and Aspergillus fumigatus.     

Metabolite profiling of fecal water extracts from human colorectal cancer

Colorectal cancer is the second leading cause of cancer death in developed countries. There is a need for better preventive strategies to improve the outcome of this disease. The increasing availability of high‐throughput methodologies opens up new possibilities for screening new markers. The application of NMR metabolic profiling to fecal water extracts has interesting potential as a diagnostic tool for detecting colorectal cancer. We obtained NMR metabolic profiles of fecal water extracts from patients with colorectal cancer and healthy individuals, to characterize possible differences between them and to identify potential diagnostic markers. Our results show that metabolic profiling of fecal water extracts is a cheap, reproducible and effective method for detecting colorectal cancer markers and therefore complements other stool‐screening methods. A low concentration of short‐chain fatty acids, such as acetate and butyrate, previously associated with the development of colorectal cancer, appears to be the most effective marker. Concentrations of proline and cysteine, which are major components of most colonic epithelium mucus glycoproteins, also display significant changes in samples from colorectal cancer. Differentiation between fecal water extracts from controls and patients with colorectal cancer by NMR spectroscopy combined with chemometric techniques opens up new possibilities for developing new, efficient, high‐throughput screening protocols. Copyright © 2008 John Wiley & Sons, Ltd.