Fecal Suspension Research Articles

Fecal micobiota transplants - The clinical perspective

Abstract: Through analytical advances, the complexity of the human microbiome has recently become apparent. The emerging data have shed light on the microorganisms living in intimate relationship with each cell in the body, and on how much of a person's daily function and health depends on this symbiosis. Fecal microbiota transplantation (FMT), also called bacteriotherapy, refers to the infusion of a fecal suspension from one individual into the gastrointestinal tract of another person, most often performed to cure diseases caused by microbial dysbiosis. FMT is by no means a new therapeutic modality, and was described as early as the fifth century. Nevertheless, until recently, only few peer-reviewed case reports could be found in the medical literature. In the past 3 years, stool has emerged as a biologically active, complex mixture of living organisms with great therapeutic potential for a number of diseases, most notably recurrent Clostridium difficile infection (CDI). Many case series and a number of clinical trials have repeatedly shown high cure rates, of up to 93%, in patients with recurrent CDI, compared with 30-40% cure obtained with standard antimicrobial treatment. FMT can be delivered via enema, colonoscopy, nasoduodenal tube, nasogastric tube and orally, as encapsulated FMT. An overview will be given about the current knowledge with regard to efficacy and safety of different delivery methods, focusing on oral therapy as the most convenient and cost-effective route. The development of the FMT capsule will be described as will our experience providing FMT to over 200 patients as part of clinical trials or routine care, including long-term follow up data. Few FMT-related adverse events have been reported to date, and most were related to the delivery mechanism. The potential use of FMT for other indications will be briefly explored, including inflammatory bowel diseases, functional bowel disorders, de-colonization of resistant pathogens and, through modification of the gut-brain axis, neuropsychiatric disorders.

Fecal Microbiota Transplantation: Current Applications, Effectiveness, and Future Perspectives.

Fecal microbiota transplantation (FMT) is the infusion of liquid filtrate feces from a healthy donor into the gut of a recipient to cure a specific disease. A fecal suspension can be administered by nasogastric or nasoduodenal tube, colonoscope, enema, or capsule. The high success rate and safety in the short term reported for recurrent Clostridium difficile infection has elevated FMT as an emerging treatment for a wide range of disorders, including Parkinson’s disease, fibromyalgia, chronic fatigue syndrome, myoclonus dystopia, multiple sclerosis, obesity, insulin resistance, metabolic syndrome, and autism. There are many unanswered questions regarding FMT, including donor selection and screening, standardized protocols, long-term safety, and regulatory issues. This article reviews the efficacy and safety of FMT used in treating a variety of diseases, methodology, criteria for donor selection and screening, and various concerns regarding FMT.

Oral vaccine of Lactococcus lactis harbouring pandemic H1N1 2009 haemagglutinin1 and nisP anchor fusion protein elevates anti-HA1 sIgA levels in mice.

An oral lactococcal-based vaccine which haboured the haemagglutinin1 (HA1) antigen fused to nisP anchor protein for the purpose of surface displaying the HA1 antigen was developed against H1N1 virus. Recombinant L. lactis strains expressed HA1-nisP fusion proteins when induced with nisin, as confirmed through western blotting. However, immunofluorescense did not detect any surface-displayed proteins, suggesting that the protein was either unsuccessfully translocated or improperly displayed. Despite this, oral administration of recombinant L. lactis strains to BALB/c mice revealed that significant levels of anti-HA1 sIgA antibodies were detected in mice fecal suspension samples of mice group NZ9000 (pNZ:HN) when compared to the negative control NZ9000 (pNZ8048) group. Specific anti-HA1 sIgA antibodies were locally produced and live recombinant lactococcal vaccine was able to elicit humoral response of BALB/c mice despite unsuccessful surface display of the HA1 epitope.

In vitroEvaluation of Plant Derived Dietary Fibers as Prebiotic for Indian Leopard (Panthera pardus fusca)

An in vitro experiment was conducted to determine the prebiotics potential of plant derived fibres (Jerusalem artichoke (JA) and chicory (CH)) and animal derived substrates (ADS) (chicken cartilage (CC) and goat tendon (GT)) as compared to positive control of fructo-oligosaccharides (FOS). Eleven Indian leopards fed meat only diets were used as donor animals. Pooled faecal samples were diluted 1: 20 (w/v), 30 ml of buffered faecal suspension and 250 mg substrate were placed into 100 ml fermentation glass syringe and incubated for 24 h. Each substrate was incubated in quadruplicate, repeated three times at weekly interval. Cumulative gas production (CGP) was measured at 0, 3, 6, 12, 18 and 24 h. The CGP, and concentrations of short chain fatty acids (SCFA) and lactate were higher (P<0.01), with simultaneous decrease (P<0.01) in concentration of ammonia, branched chain fatty acids and pH when plant derived substrtaes (PDS) were incubated as compared to ADS. It is evident that the PDS are better fermentation substrate than ADS. Beneficial effects of using PDS as substrates included decreased pH and ammonia level, and increased concentration of butyrate and lactate. Between the PDS, JA showed the best response in terms of reduction of pH, ammonia and improvement in the concentration of butyrate and lactate. It was concluded that JA has the potential to be used as prebiotic in the diet of captive Indian leopard fed meat only diets.

In Vitro Effects of Thymol-β-d-Glucopyranoside on Salmonella enterica Serovar Typhimurium and Escherichia coli K88

Although thymol is bactericidal against many pathogens in vitro, its in vivo effectiveness against pathogens in the lower gastrointestinal tract is limited because of its rapid absorption in the proximal gut. Thymol-β-<small>D</small>-glucopyranoside (β-thymol), a conjugated form of thymol, can deliver thymol to the lower gastrointestinal tract and has shown antibacterial effects. In the present study, we examined the in vitro effects of β-thymol on Salmonella enterica serovar Typhimurium (ST) and Escherichia coli K88 (K88). We inoculated one-half strength Mueller-Hinton broth with 5.8 ± 0.09 log CFU/ml novobiocin- and naladixic acid-resistant (NN) ST (NVSL 95-1776) and 5.1 ± 0.09 log CFU ml(-1) NN-resistant K88, with or without porcine feces (0.1% [wt/vol]) (fecal incubations). The resultant bacterial suspensions were distributed under N2 to triplicate sets of tubes to achieve initial concentrations of 0, 3, 6, and 12 mM for ST treatments and 0, 3, 12, and 30 mM for K88 treatments. Samples were incubated at 39°C and then plated onto NN-containing brilliant green agar and NN-containing MacConkey agar; ST and K88 CFU concentrations were determined via 10-fold dilutions, and viable cell counts were performed at 0, 6, and 24 h. No differences in ST CFU counts were observed in β-thymol-treated tubes without the added porcine feces (i.e., pure culture) at 6 or 24 h. However, in tubes that contained fecal incubations, ST CFU counts were reduced (P < 0.05) from controls at 6 h in tubes treated with 6 and 12 mM β-thymol, whereas in tubes treated with 3, 6, and 12 mM β-thymol the CFU counts were reduced (P < 0.05) at 24 h. No differences were observed in K88 CFU counts in pure culture or in fecal incubations at 6 h, but K88 CFU counts were reduced (P < 0.05) in both pure and fecal incubations at 24 h. The results from this study demonstrate that β-thymol, in the presence of fecal suspensions, has anti-Salmonella and anti-E. coli effects, suggesting a role of β-glycoside-hydrolyzing microbes for the release of bactericidal thymol from β-thymol.

G and P types of bovine group A rotavirus in northern India

Animal rotaviruses (RVs) are considered as a potential threat to humans due to possibility of interspecies transmission and exchange of genomic materials. Among several RVs, the group A rotaviruses (RVA) are the major cause of diarrhoea in cattle and buffalo calves worldwide. The present study was carried out to understand epidemiology and types of RVA circulating in Mathura region of northern India. One hundred faecal samples were collected from diarrhoeic cattle (n=94) and buffalo calves (n=6) from organized dairy farms. Viral RNA was extracted from faecal suspension and was transcribed to cDNA using RT-PCR. The cDNA was amplified for VP6, VP7 and VP4 genes and multiplex nested PCR was done for G and P genotyping. Twelve samples were detected positive for RVA by antigen detection ELISA and eleven samples produced expected amplicon for group A specific VP6 gene. On genotyping with G3, G6, G8 and G10 specific primers for VP7 gene and P [1] and P [11] specific primers for VP4 gene, G6 genotype predominated over other genotypes. Dual genotypes were also observed. In P typing only one isolate was found to be positive for P [1] and rest all samples remained untypable. The result of present study indicates G6 as a major G genotype and change in frequencies of distribution of G types in this part of the country.

Comparison of fecal pooling methods and DNA extraction kits for the detection of Mycobacterium avium subspecies paratuberculosis

The aim of the study was to develop a sensitive method using quantitative real-time polymerase chain reaction (qPCR) with pooled fecal samples for the screening of Johne's disease (JD). Manufacturer-specified and our new pooling method in combination with five commercial kits for DNA extraction and purification were compared. Different volumes of pooled fecal suspensions were tested, and the results were compared for individual samples and three pool sizes (5, 10, and 50 samples); each of the fecal suspensions, which were prepared from healthy dairy and beef cattle was spiked with 0, 10, 100, or 1000 cultured Mycobacterium avium subspecies paratuberculosis (MAP) organisms or was mixed with fecal suspensions from experimentally infected cattle. The MAP DNA detection proportion with our pooling method in combination with Johne-Spin kit (Fasmac, Japan) was 100% for all models and all pool sizes, except for the low shedder model with a pool size of 50. There was no loss of sensitivity in pools of 10 subjects or less by using the new method. These results suggest that new method is a sensitive, practical, and cost-effective screening test for the detection of MAP-infected cattle and the monitoring of JD-free herds.

Beating Cancer with a Gut Feeling.

Blockade of immune checkpoint molecules, a group of molecules normally involved in maintaining self-tolerance and limiting T cell responses, has emerged as a breakthrough in cancer therapy. Two recent studies published in Science show that, in mice, gut commensal microbes promote antitumor immunity and may determine therapy efficacy.

Fecal Microbiota Transplantation: Harnessing the Gut Microbiota to Treat Disease

Fecal microbiota transplantation (FMT) involves the transfer of consortia of bacteria from healthy individuals to patients with Clostridium difficile infection (CDI) or other conditions to promote recolonization with missing components of intestinal flora. FMT dates as far back as 4th-century China when human fecal suspensions were used to treat food poisoning and severe diarrhea and has been used in veterinary medicine to treat ruminal disorders at least as far back as the 17th century.

Fecal microbiota transplantation in gastrointestinal diseases: what practicing physicians should know.

Clostridium difficile infection (CDI) is one of the most commonly reported nosocomial pathogens in the United States and Europe, with recent CDI-associated mortality in the United States approaching 30 000 deaths annually. Antibiotics remain the preferred treatment for CDI; however, a minority of patients experience numerous relapses and are treated with restoration of the bowel microbiota, termed fecal microbiota transplantation (FMT). FMT involves the introduction of a fecal suspension from a healthy donor into the gut of the infected patient to cure the CDI and replace depleted components of the gut microbiota. FMT is particularly effective and safe in curing CDI, using a colonoscope or enema to deliver 1 to 2 infusions. Given that 6425 CDIs were reported in Poland in 2014, practicing physicians should understand the benefits and limitations of FMT in CDI as this novel therapy has rapidly advanced to the level of the "standard-of-care" status in Australia, the United States, and many parts of Europe. FMT has been administered either as a suspension in saline, a highly refined liquid product which can be frozen, as lyophilized powder in capsules, and as an encapsulated spore preparation. The ultimate products to reach the market will be shaped by the indications approved by regulatory bodies. At present, the fecal suspension in saline remains the treatment of choice to terminate relapsing and severe CDI, which we will review here. The use of FMT for non-CDI indications, such as inflammatory bowel disease and irritable bowel syndrome, is likely to increase. At present, these indications remain in the domain of research institutions.

Fast Technology Analysis Enables Identification of Species and Genotypes of Latent Microsporidia Infections in Healthy Native Cameroonians.

Several enteric microsporidia species have been detected in humans and other vertebrates and their identifications at the genotype level are currently being elucidated. As advanced methods, reagents, and disposal kits for detecting and identifying pathogens become commercially available, it is important to test them in settings other than in laboratories with "state-of-the-art" equipment and well-trained staff members. In the present study, we sought to detect microsporidia DNA preserved and extracted from FTA (fast technology analysis) cards spotted with human fecal suspensions obtained from Cameroonian volunteers living in the capital city of Yaoundé to preclude the need for employing spore-concentrating protocols. Further, we tested whether amplicon nucleotide sequencing approaches could be used on small aliquots taken from the cards to elucidate the diversity of microsporidia species and strains infecting native residents. Of 196 samples analyzed, 12 (6.1%) were positive for microsporidia DNA; Enterocytozoon bieneusi (Type IV and KIN-1), Encephalitozoon cuniculi, and Encephalitozoon intestinalis were identified. These data demonstrate the utility of the FTA cards in identifying genotypes of microsporidia DNA in human fecal samples that may be applied to field testing for prevalence studies.

Structural Transformation of 8-5-Coupled Dehydrodiferulates by Human Intestinal Microbiota.

Ingested dehydrodiferulates (DFAs) are partially released from cereal dietary fiber by human colonic microbiota, but little research has explored the further microbial metabolism of 8-5-coupled DFAs. This study investigated the in vitro microbial metabolism and elucidated major metabolites of free 8-5-DFAs (benzofuran and open forms) and an esterified analogue, 8-5-DFA diethyl ester (benzofuran). Synthesized standard compounds were incubated with fresh human fecal suspensions. Metabolites were isolated and structurally elucidated using high-resolution-LC-time-of-flight-(ToF)-MS, GC-MS, and NMR. Nine metabolite structures were unambiguously characterized with NMR, and four additional metabolites were tentatively identified to reveal structural conversion motifs: propenyl side chain hydrogenation (all substrates), O-demethylation and reductive ring-opening (8-5-DFA diethyl ester and free 8-5-DFA [benzofuran]), and de-esterification (8-5-DFA diethyl ester). A pathway of microbial 8-5-DFA metabolism was proposed based on metabolite formation kinetics. Importantly, de-esterification of the 8-5-DFA diethyl ester occurred primarily after and/or concurrently with other metabolism steps. Cleavage to monomers was not observed.

Therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis

Objective To investigate the therapeutic effects and the possible mechanism of fecal transplantation on rats with Clostridium difficile-associated pseudomembranous colitis. Methods A total of 40 Sprague-Dawley rats were divided into four groups including the healthy control group, model group, fecal transplant treatment group and vancomycin treatment group. Rats in three experimental groups were subcutaneously injected with clindamycin phosphate (10 mg), followed by treatment with toxin producing Clostridium difficile (ACTT43255) enema 24 hours later. The rats in fecal transplant treatment group and vancomycin treatment group were respectively treated with fecal suspension and vancomycin one day after modeling. The rats were fasted for one day after the last administration and then executed. The levels of potassium ion (K), sodium ion (Na), albumin (ALB), white blood cells (WBC), C-reaction protein (CRP), interleukin-1β (IL-1β), interleukin-10 (IL-10), interleukin-12 (IL-12) and interleukin-17 (IL-17) as well as the percentage of neutrophils (N%) in serum samples were detected. The colon tissue samples were collected for pathology examination. Results The rat model of pseudomembranous colitis was successfully established by subcutaneous injection of clindamycin in combination with toxin-producing Clostridium difficile (ACTT43255) enema. The signs of intestinal inflammation including serious weight loss, remarkably shortened colon length and significantly increased colon wet weight index were observed in rats from the model group (P<0.05). Compared with the rats from model group, the rats received fecal transplant showed significantly increased levels of K, ALB, IL-10 and IL-10/IL-12 in serum and decreased levels of WBC, N%, CRP, IL-1β and IL-17 (P<0.05). Conclusion Fecal transplantation was proved to be an effective approach for the treatment of pseudomembranous colitis. The therapeutic mechanism might due to its impacts on serum inflammatory factors. Key words: Fecal transplantation; Pseudomembranous colitis; Inflammatory factor

Stability of Individual Maillard Reaction Products in the Presence of the Human Colonic Microbiota.

Maillard reaction products (MRPs) are taken up in substantial amounts with the daily diet, but the majority are not transported across the intestinal epithelium. The aim of this study was to obtain first insights into the stability of dietary MRPs in the presence of the intestinal microbiota. Four individual MRPs, namely, N-ε-fructosyllysine (FL), N-ε-carboxymethyllysine (CML), pyrraline (PYR), and maltosine (MAL), were anaerobically incubated with fecal suspensions from eight human volunteers at 37 °C for up to 72 h. The stability of the MRPs was measured by HPLC with UV and MS/MS detections. The Amadori product FL could no longer be detected after 4 h of incubation. Marked interindividual differences were observed for CML metabolism: Depending on the individual, at least 40.7 ± 1.5% of CML was degraded after 24 h of incubation, and the subjects could thus be tentatively grouped into fast and slow metabolizers of this compound. PYR was degraded by 20.3 ± 4.4% during 24 h by all subjects. The concentration of MAL was not significantly lowered in the presence of fecal suspensions. In no case could metabolites be identified and quantified by different mass spectrometric techniques. This is the first study showing that the human colonic microbiota is able to degrade selected glycated amino acids and possibly use them as a source of energy, carbon, and/or nitrogen.

Development and Validation of an in vitro Experimental GastroIntestinal Dialysis Model with Colon Phase to Study the Availability and Colonic Metabolisation of Polyphenolic Compounds.

The biological effects of polyphenols depend on their mechanism of action in the body. This is affected by bioconversion by colon microbiota and absorption of colonic metabolites. We developed and validated an in vitro continuous flow dialysis model with colon phase (GastroIntestinal dialysis model with colon phase) to study the gastrointestinal metabolism and absorption of phenolic food constituents. Chlorogenic acid was used as model compound. The physiological conditions during gastrointestinal digestion were mimicked. A continuous flow dialysis system simulated the one-way absorption by passive diffusion from lumen to mucosa. The colon phase was developed using pooled faecal suspensions. Several methodological aspects including implementation of an anaerobic environment, adapted Wilkins Chalgren broth medium, 1.10(8) CFU/mL bacteria suspension as inoculum, pH adaptation to 5.8 and implementation of the dialysis system were conducted. Validation of the GastroIntestinal dialysis model with colon phase system showed a good recovery and precision (CV < 16 %). Availability of chlorogenic acid in the small intestinal phase (37 ± 3 %) of the GastroIntestinal dialysis model with colon phase is comparable with in vivo studies on ileostomy patients. In the colon phase, the human faecal microbiota deconjugated chlorogenic acid to caffeic acid, 3,4-dihydroxyphenyl propionic acid, 4-hydroxybenzoic acid, 3- or 4-hydroxyphenyl acetic acid, 2-methoxy-4-methylphenol and 3-phenylpropionic acid. The GastroIntestinal dialysis model with colon phase is a new, reliable gastrointestinal simulation system. It permits a fast and easy way to predict the availability of complex secondary metabolites, and to detect metabolites in an early stage after digestion. Isolation and identification of these metabolites may be used as references for in vivo bioavailability experiments and for investigating their bioactivity in in vitro experiments.

PTH-228 Genotyping of rotavirus isolates prior to the introduction of the rotavirus vaccine in scotland and early indications of the impact of the vaccine

<h3>Introduction</h3> Rotaviruses (RV) are the leading cause of severe gastroenteritis in children less than five years of age worldwide. Rotarix®, a live attenuated monovalent vaccine containing a RV strain of G1P[8] specificity has been included in the routine childhood immunisation schedule from June 2013 in Scotland and is offered to infants under the age of four months in two doses. This study aimed to characterise the prevalent rotavirus strains in Scotland prior to the introduction of the vaccine. <h3>Method</h3> Rotavirus positive faecal samples from various Scottish regional virology laboratories covering the years 2012–2013, before the introduction of the vaccine, were genotyped. Viral RNA was extracted from faecal suspensions using guanidine isothiocyanate – silica gel extraction method. VP7 and VP4 gene specific multiplex hemi-nested PCRs were used for genotyping. Surveillance systems were established by Health Protection Scotland to monitor the impact of the vaccine. <h3>Results</h3> A total of 387 samples were genotyped from the regional virology laboratories in Aberdeen, Dundee, Inverness, Glasgow and Edinburgh. The commonest strain in Scotland was G1P[8] accounting for 72.1% of cases. The other strains that were identified were G2P[4] in 7.2%, G4P[8] in 6.9%, G9P[8] in 3.4% and G3P[8] in a further 2.3% of cases. Single cases of G12P[8] and G9P[4] were also found (0.26% respectively). Mixed infections were seen in 5.4% of cases. The provisional uptake for the first dose of Rotarix® vaccine in Scotland for children born between 1^stof July to 30^thSept 2013 was 92.7%% for the completed course by 12 months. The laboratory confirmed cases of RV reported to HPS via ECOSS (The Electronic Communication of Surveillance in Scotland) showed a marked reduction in confirmed reports of RV compared to a three year average of the years 2011–13. Surrogate markers, e.g. calls to NHS24 and GP consultations with childhood diarrhoea and vomiting, have also shown a downward trend. <h3>Conclusion</h3> The pre-vaccine surveillance of RV strains confirms that G1P[8] is the predominant strain in Scotland. There has been a promising reduction in laboratory confirmed cases of RV in Scotland following the introduction of the vaccine. The baseline genotyping data will be used to ascertain cross protection against strains and also identify vaccine induced RV strain shifts and an overall evaluation of the programme. <h3>Disclosure of interest</h3> None Declared.

Toxoplasma gondii B1 Gene Detection in Feces of Stray Cats around Seoul, Korea and Genotype Analysis of Two Laboratory-Passaged Isolates.

The increasing prevalence of Toxoplasma gondii infection in the human population in the Republic of Korea (= Korea) is due to various reasons such as an increase in meat consumption. However, the importance of cats in transmitting T. gondii infection through oocysts to humans has seldom been assessed. A total of 300 fecal samples of stray cats captured around Seoul from June to August 2013 were examined for T. gondii B1 gene (indicating the presence of oocysts) using nested-PCR. Fourteen (4.7%) of 300 cats examined were positive for B1 gene. Female cats (7.5%) showed a higher prevalence than male cats (1.4%). Cats younger than 3 months (5.5%) showed a higher prevalence than cats (1.5%) older than 3 months. For laboratory passage of the positive samples, the fecal suspension (0.2 ml) of B1 gene positive cats was orally inoculated into experimental mice. Brain tissues of the mice were obtained after 40 days and examined for the presence of tissue cysts. Two isolates were successfully passaged (designated KNIH-1 and KNIH-2) and were molecularly analyzed using the SAG5D and SAG5E gene sequences. The SAG5D and SAG5E gene sequences showed high homologies with the ME49 strain (less virulent strain). The results indicated the importance of stray cats in transmitting T. gondii to humans in Korea, as revealed by detection of B1 gene in fecal samples. T. gondii isolates from cats were successfully passaged in the laboratory for the first time in Korea.

Destruction of the Capsid and Genome of GII.4 Human Norovirus Occurs during Exposure to Metal Alloys Containing Copper.

Human norovirus (HuNoV) represents a significant public health burden worldwide and can be environmentally transmitted. Copper surfaces have been shown to inactivate the cultivable surrogate murine norovirus, but no such data exist for HuNoV. The purpose of this study was to characterize the destruction of GII.4 HuNoV and virus-like particles (VLPs) during exposure to copper alloy surfaces. Fecal suspensions positive for a GII.4 HuNoV outbreak strain or GII.4 VLPs were exposed to copper alloys or stainless steel for 0 to 240 min and recovered by elution. HuNoV genome integrity was assessed by reverse transcription-quantitative PCR (RT-qPCR) (without RNase treatment), and capsid integrity was assessed by RT-qPCR (with RNase treatment), transmission electron microscopy (TEM), SDS-PAGE/Western blot analysis, and a histo-blood group antigen (HBGA) binding assay. Exposure of fecal suspensions to pure copper for 60 min reduced the GII.4 HuNoV RNA copy number by ∼3 log10 units when analyzed by RT-qPCR without RNase treatment and by 4 log10 units when a prior RNase treatment was used. The rate of reduction of the HuNoV RNA copy number was approximately proportional to the percentage of copper in each alloy. Exposure of GII.4 HuNoV VLPs to pure-copper surfaces resulted in noticeable aggregation and destruction within 240 min, an 80% reduction in the VP1 major capsid protein band intensity in 15 min, and a near-complete loss of HBGA receptor binding within 8 min. In all experiments, HuNoV remained stable on stainless steel. These results suggest that copper surfaces destroy HuNoV and may be useful in preventing environmental transmission of the virus in at-risk settings.

Duodenal Application Is the Method of Choice

We would like to thank the authors for their insightful review of Clostridium difficile infection (CDI) (1) and to add the following information to complement the section about stool transplantation: At the SRH Kurpfalz Hospital, 22 stool transplantations (allogenic intestinal recolonizations) have been performed to treat CDI to date. In all of these cases, donor feces was infused exclusively into the duodenum using an endoscope, in line with the method described by van Nood (2). The clinical response rate was comparable with published data; 16 of 17 patients were relapse-free during the 3-month follow-up period. Due to the pooling of the data on gastric and duodenal administration, no inferiority of the antegrade versus the retrograde route of administration can be derived from Table 3. In contrast to the information provided in the table, the 18 patients of Aas et al. (3) received the fecal suspension by gastric delivery. Thus, in 3 of 6 studies (45 of 101 patients) gastric application of the fecal suspension was performed, involving the risk of aspiration pneumonia by fecal bacteria, besides the inactivation of the donor microbiota by gastric acid. Another significant difference in the methods used is the preceding bowel lavage which, among the studies with antegrade application, was only used in the study by van Nood which was also the only randomized controlled trial (2). Colonoscopic application is more taxing for the patient and associated with the risk of perforation of the inflamed colon. Provided the patients receive sedation with propofol, the acceptance is excellent. The passage of the donor microbiota through the small intestine does not represent an undesirable contamination but is rather seen as favorable, as it accelerates the reconstitution of the resident microbiota. Thus, we consider duodenal application after peroral bowl lavage as described by van Nood as the reference method until other data from randomized trials have become available.

EFFECT OF SILVER NANOPARTICLES IN EXPERIMENTAL PERITONITIS

In our research we have investigated the effect of intraperitoneal introduction of silver nanoparticles solution (linear particle size of 10–20 nm) into intact animals and in the simulation of experimental peritonitis. We have evaluated the indices of nonspecific proteinases and their inhibitors in blood serum and peritoneum lavage. The intraperitoneal introduction of silver nanoparticles solution to intact animals leads to the minimal reaction of proteinase-inhibitor systems components in blood serum and peritoneum lavage. When modeling inflammatory process in peritoneum cavity by intraperitoneal injection of 10% filtered fecal suspension of rats, application of silver nanoparticles solution is accompanied by decrease in extent of activation of proteinases and preservation of inhibitory potential, both at systemic and local level, which can indicate antiinflammatory effects of nanosilver.