Fecal Suspension Research Articles

Successful Treatment of Recurrent Clostridium difficile Infection with Donor Stool Administered at Colonoscopy: A Case Series

Purpose: Chronic relapsing Clostridium difficile infections (CDI) are increasingly common and challenging to manage. Current guidelines recommend a tapering course of vancomycin after a second recurrence, however up to 60% of these patients develop further recurrence after vancomycin is stopped. Fecal bacteriotherapy involves administration of feces from a healthy individual to promote recolonization with missing components of gut flora. We report our experience treating 12 patients with relapsing CDI using a suspension of donor stool administered through the colonoscope. Methods: Patients were asked to identify a donor (partner or family member) who had not taken antibiotics within the past 90 days. Donor stool was tested for bacterial culture, Giardia antigen and C. difficile toxins A & B. Patient and donor were tested for HIV 1 & 2, hepatitis A, B, & C, and syphilis. Donors took a dose of milk of magnesia the night prior to the procedure (to facilitate optimum consistency for processing) and submitted a fresh stool specimen on the day of the procedure. Patients discontinued vancomycin 3 days prior to the procedure and performed a standard PEG bowel preparation. Immediately before colonoscopy, 6 to 8 tablespoons of donor stool was added to 1 liter of sterile water and shaken vigorously to homogenize. Colonoscopy was performed, and the suspension of donor stool in sterile water was delivered through the colonoscope in 60 cc aliquots starting the terminal ileum (or cecum). Patients were asked to avoid defecating for at least 45 minutes after the procedure. Follow up included telephone contacts and as needed visits to the practice. Results: 12 patients (11 female, 1 male), with mean age of 55.6 (range 19-80) and mean CDI duration of 16.8 months (range 3-84 months) were treated using the above protocol. All had relapsed after metronidazole, rifaximin, Saccharomyces boulardii and repeated tapering courses of vancomycin or were taking vancomycin chronically. Two had been treated with intravenous immunoglobulin. An average of 740 cc (range 500-960 cc) of fecal suspension was delivered. All patients tolerated the procedure well. None have had a documented recurrence of CDI to date with a mean follow up period of 7.4 months (range 2-19 months). Ten have remained symptom free. Two had diarrhea after the procedure, but both were C. difficile negative. One responded to treatment with a fiber supplement and the other resumed vancomycin. Conclusion: Fecal bacteriotherapy administered at colonoscopy is an effective, safe and well tolerated option for patients with chronic relapsing CDI refractory to other therapies. We provide a simple and easily reproducible protocol for performing this treatment.

Durable Alteration of the Colonic Microbiota by the Administration of Donor Fecal Flora

To determine whether fecal bacteriotherapy results in a durable beneficial change in the colonic microbiota of patients with flora-related disorders. Earlier studies have implicated the colonic microbiota in a number of conditions. Administration of a fecal suspension from a healthy individual to an ill individual (fecal bacteriotherapy) can cure Clostridium difficile infection and potentially other diseases. Oral probiotics do not work in this condition, yet there has been no study to determine whether fecal bacteriotherapy results in prolonged implantation. Fecal samples were collected from 10 patients undergoing fecal bacteriotherapy. Patients completed an antibiotic schedule and bowel lavage before the infusion of healthy donor feces. Using a molecular approach, the bacterial populations in patient fecal samples were followed from pretreatment to 24 weeks post-initial infusion and compared with the initial infused donor fecal suspension. At intervals of 4, 8, and 24 weeks after the procedure, the bacterial populations in the patients' fecal samples consisted predominantly of bacteria derived from the healthy donor samples. Comparisons of similarity at 4, 8, and 24 week samples to the donor-infused sample were made and each recipient's baseline sample was statistically significant with Friedman test. This study demonstrates a durable beneficial change in the patients' bacterial populations of the colon to represent those of the healthy donor's microbiota. Manipulation of the colonic microbiota to improve its protective and beneficial role represents a promising field of new therapeutic strategies for the treatment of gastrointestinal conditions.

Is the abundance of Faecalibacterium prausnitzii relevant to Crohn's disease?

Reports that bacteria within the Firmicutes phylum, especially the species Faecalibacterium prausnitzii, are less abundant in Crohn's disease (CD) patients and supernatants from cultures of this bacterium are anti-inflammatory prompted the investigation of the possible correlations between the abundance of F. prausnitzii and the response to treatment in patients with gut diseases and healthy controls. In a randomized, double-blind trial, faeces were collected from healthy volunteers, and from patients with active CD, ulcerative colitis (UC) and irritable bowel syndrome before and after treatment. The levels of F. prausnitzii DNA in faecal suspensions were determined by PCR. Treatment by an elemental diet was effective, resulting in decreases in both the Harvey and Bradshaw index (P<0.001) and the concentrations of serum C-reactive protein (P<0.05). The total levels of F. prausnitzii in faecal samples from CD patients at presentation were lower than those in the other groups both before and after the treatment. There was no correlation between F. prausnitzii abundance and the severity of CD before treatment. Clinical improvement unexpectedly correlated with a significant decrease in the abundance of F. prausnitzii, especially the A2-165 subgroup (P<0.05). Our data suggest that a paucity of F. prausnitzii in the gastrointestinal microbial communities is likely to be a minor aetiological factor in CD: recovery following elemental diet is attributed to lower levels of gut flora.

Norovirus Gastroenteritis Outbreak is Associated with Contaminated Drinking Water in South China

Noroviruses (NoVs) cause gastroenteritis in all age groups. To investigate an acute gastroenteritis outbreak in Shenzhen, South China, we analyzed the presence of NoVs in fecal specimens and drinking water samples. Viral RNAs were extracted from fecal suspensions and water concentrates made by membrane adsorption-elution, and NoV was detected by reverse transcription-polymerase chain reaction (RT-PCR). Among the 43 diseased workers at the two factories and one construction site, NoVs were detected in 19 out of 28 stool specimens (68%) and 3 out of 16 water samples (19%). Importantly, the six NoV strains identified from the water and the stool samples share over 99% sequence identity and all belong to GII 4 viruses. Epidemiological survey also identified a water pipe breakage possibly causing the water contamination, and confirmed that the contaminated drinking water largely contributed to the NoV gastroenteritis outbreak. To our knowledge, this is the first time that a viral concentration and detection method for waterborne NoV is applied to a gastroenteritis outbreak in China.

Lactoferrin increases both resistance to Salmonella typhimurium infection and the production of antibodies in mice

Lactoferrin (Lf) is a multifunctional iron-binding glycoprotein with antibacterial and immunomodulatory activities. The antibacterial influence of orally administered bovine Lf (bLf) against murine infection caused by Salmonella typhimurium ( S. typhimurium) has scarcely been explored. In the current study, Balb/c mice were treated orally for 7 days with either 5 or 100 mg of bovine lactoferrin (bLf). On day 7 of treatment, mice were intragastrically infected with a lethal or sublethal dose of colony forming units (CFU) of S. typhimurium. During treatment with bLf, feces from mice sublethally infected were harvested daily to prepare fecal suspensions, which were serially diluted and plated onto Salmonella Shigella agar to estimate CFU/g of feces. After sacrificing the animals on day 7, 14 or 21 post-infection, samples of intestinal fluid, Peyer's patches (PP), liver and spleen were collected to count the number of CFU by plate dilution. Intestinal secretions were also employed, along with serum samples, to evaluate total IgA, IgG and IgM antibodies, and those against Salmonella surface proteins and bLf by ELISA assay. In lethally infected mice both bLf doses decreased mortality. In sublethally infected mice, both bLf doses decreased bacterial shedding in feces and intestinal fluid, and also reduced bacterial colonization at PP and bacterial translocation in the liver and spleen. Levels of total and those IgG and IgM in serum and IgA in intestinal secretions against Salmonella surface proteins and bLf were enhanced with both doses of bLf. These findings suggest that the effect of bLf against the infection by S. typhimurium in mice may be the result of an antimicrobial activity linked with its modulatory effect on immunocompetent cells (from intestinal and peripheral organs) involved in antibody production.

Reação de hemi-nested RT-PCR dirigida ao gene da hemaglutinina-esterase do Coronavírus Bovino

O coronavírus bovino (BCoV) é um vírus RNA simples fita, de sentido positivo, não segmentado com envelope constituído de uma camada dupla de lipídios com quatro proteínas (HE, S, E e M) que resultam no aspecto de coroa dos vírions. Como a HE (hemaglutinina-esterase) é uma proteína polimórfica com uma função de receptor aglutinante secundária, estudos sobre a diversidade do gene HE podem possibilitar maiores informações sobre a evolução e interação hospedeiro-parasita do BCoV. Uma reação de hemi-nested RT-PCR foi desenvolvida para a amplificação de um produto de 441pb do gene HE do BCoV (nt 543 ao 562). Temperaturas ótimas de hibridização foram testadas em um termociclador com gradiente para a reação de hemi-nested e utilizada no protocolo final. A sensibilidade analítica foi determinada por meio da diluição serial na base 10 do BCoV amostra Kakegawa (título HA: 256) em uma suspensão fecal negativa para BCoV, resultando positiva até a diluição de 10-6, mostrando uma alta sensibilidade analítica sem inibição na PCR. O protocolo final da hemi-nested RT-PCR foi aplicado a 21 amostras fecais de vacas previamente positivas para BCoV e o sequenciamento de DNA do produto de 441pb de 14 amostras resultaram em sequências com elevado escore do gene HE do BCoV após a análise no BLAST/n. Essa hemi-nested RT-PCR é uma ferramenta poderosa para estudos de diversidade filogenética de linhagens de campo de BCoV e para estudos comparativos entre os diferentes genes dos Coronavírus.

Erythropoietin improves skeletal muscle microcirculation in mice with sepsis through activation of eNOS

This study is to explore the potential mechanisms involved in the improvement of microcirculation by the EPO in septic mice.MethodsSepsis was induced in mice by intraperitoneal injection of a fecal suspension (12.5 mg/mouse). The septic mice were given rHuEPO (400 U/kg) 18 hrs after sepsis. Capillary perfusion and nicotinamide adenine dinucleotide (NADH) fluorescence were observed using an intravital microscopy. In addition, endothelial cells derived from the skeletal muscle were treated with rHuEPO (5 U/ml) and eNOS activation and activity were assessed.ResultsSeptic mice demonstrated microcirculatory dysfunction with a decreased capillary perfusion and an increased tissue NADH fluorescence indicating impaired tissue bioenergetics. In contrast, septic mice treated with rHuEPO resulted in an improvement in the perfused capillary density and decreased muscle NADH fluorescence. However, this beneficial effect of rHuEPO did not occur in septic mice treated with L‐NAME (20 mg/kg) or mice deficient in eNOS. Treatment of endothelial cells with rHuEPO resulted in an increase in eNOS phosphorylation and NO production.In conclusionOur results suggest that eNOS plays an important role in mediating the beneficial effect of rHuEPO on microcirculation in this model of sepsis. (Supported by funding from Department of National Defense, Canadian Forces Medical Group).

Liquid chromatography/mass spectrometry-based structural analysis of soyasaponin Ab metabolites by human fecal microflora

Soyasaponin Ab, a major constituent of soybean by human intestinal microflora, was anaerobically incubated with fecal suspensions from ten individuals for 48 h and its metabolites were measured by LC-MS/MS analysis. Ten metabolites were detected. The spectra of the parental constituent soyasaponin Ab showed a peak at m/z 1435 [M-H](-) ion and those of its nine metabolites showed peaks at m/z 1310.0 [M-3C(2)H(2)O-H](-) ion, m/z 1267.9 [M-4C(2)H(2)O-H](-) ion, m/z 1105.3 [M-Glc-4C(2)H(2)O-H](-) ion, m/z 973.2 [M-Glc-Ara-4C(2)H(2)O-H](-) ion, m/z 943.4 [M-2Glc-4C(2)H(2)O-H](-) ion, m/z 811.1 [M-2Glc-Ara-4C(2)H(2)O-H](-) ion, m/z 781.2 [M-2Glc-Gal-4C(2)H(2)O-H](-) ion, m/z 649.0 [M-2Glc-Gal-Ara-4C(2)H(2)O-H](-) ion, and m/z 458.8 [Soyasapogenol A+H](+) ion. Metabolic activity varied significantly between individuals. The metabolic pathway was classified into two groups: the first group potently produced soyasapogenol A and the second group accumulated soyasapogenol A 3-beta-D-glucuronide.

Simplified method to measure glucocorticoid metabolites in faeces of horses

Glucocorticoids or their metabolites can be measured in several body fluids or excreta, including plasma, saliva, urine and faeces. In recent years the measurement of glucocorticoid metabolites (GCMs) in faeces has gained increasing attention, because of its suitability for wild populations. In horses, however, the group-specific enzyme immunoassay described so far has a limited practicability due to its complex extraction procedure. Therefore, we tested the applicability of other enzyme immunoassays for glucocorticoid metabolites. The present study clearly proved that an enzyme immunoassay (EIA) for 11-oxoaetiocholanolone using 11-oxoaetiocholanolone-17-CMO: BSA (3alpha,11-oxo-A EIA) as antigen showed high amounts of immunoreactive substances. Therefore it was possible to use just a small amount of the supernatant of a methanolic suspension of faeces. The results correlated well with the already described method for measuring GCMs in horse faeces, i.e. analysing the samples with an EIA after a two step clean up procedure of the samples (Merl et al. 2000). In addition, the 3alpha,11-oxo-A EIA has the advantage of providing a bigger difference between baseline values and peak values after ACTH stimulation. The new assay increased the accuracy of the test, lowered the expenses per sample, and storing samples at room temperature after collection was less critical than with other assays investigated in our study. This is a big advantage both in the field of wildlife management of equids and in the field of equestrian sports and it shows the importance of choosing an assay which is in good accordance with the metabolites excreted in a given species.

Cell culture system for hepatitis E virus

Early studies reported propagation of hepatitis E virus (HEV) in primary hepatocytes or several established cell lines, but replication was inefficient. Recently, using inocula comprised of fecal suspensions with high loads of HEV, originally obtained from Japanese patients who contracted domestic infection of genotype 3 HEV (the JE03-1760F strain, 2.0 x 10(7) copies/ml) or genotype 4 HEV (the HE-JF5/15F strain, 1.3 x 10(7) copies/ml), we developed an efficient cell culture system for HEV in PLC/PRF/5 and A549 cells, which yielded the highest HEV load of 10(8) copies/ml in the culture supernatant, and we successfully propagated six or more generations in serial passages of culture supernatant. In addition, we constructed a full-length infectious cDNA clone (pJE03-1760F/wt) of the JE03-1760F strain, which can replicate efficiently in PLC/PRF/5 and A549 cells. Using a derivative ORF3-deficient (delta ORF3) mutant, we demonstrated that the ORF3 protein of HEV is responsible for virion egress from infected cells and is present on the surface of released HEV particles, which is associated with lipids. Various HEV strains in blood circulation were also propagated efficiently in PLC/PRF/5 and A549 cells. Our in vitro cell culture system can be used for propagation of a wide variety of HEV strains in feces and sera from various infected patients, allowing extended studies on viral replication specific to different HEV strains.

Determination of the 5′-terminal sequence of subgenomic RNA of hepatitis E virus strains in cultured cells

Using RNA preparations extracted from PLC/PRF/5 cells transfected with infectious genotype 3 hepatitis E virus (HEV) cDNA clones or inoculated with a fecal suspension containing a genotype 4 HEV, the 5'-terminal sequence of a 2.2-kb subgenomic RNA of genotype 3 and 4 HEVs was determined. Despite an insertion of T after nucleotide 5116 or an ORF3-null mutation in genotype 4 HEV and/or one of the genotype 3 variants, it was found that the subgenomic RNA of genotype 3 and 4 HEVs initiates exclusively at nucleotide 5122 with the common sequence of 5'-GC, which is identical to that of the prototype genotype 1 HEV.

Rapid detection of bovine coronavirus by a semi-nested RT-PCR

Bovine coronavirus (BCoV) is a member of the group 2 of the Coronavirus (Nidovirales: Coronaviridae) and the causative agent of enteritis in both calves and adult bovine, as well as respiratory disease in calves. The present study aimed to develop a semi-nested RT-PCR for the detection of BCoV based on representative up-to-date sequences of the nucleocapsid gene, a conserved region of coronavirus genome. Three primers were designed, the first round with a 463bp and the second (semi-nested) with a 306bp predicted fragment. The analytical sensitivity was determined by 10-fold serial dilutions of the BCoV Kakegawa strain (HA titre: 256) in DEPC treated ultra-pure water, in fetal bovine serum (FBS) and in a BCoV-free fecal suspension, when positive results were found up to the 10-2, 10-3 and 10-7 dilutions, respectively, which suggests that the total amount of RNA in the sample influence the precipitation of pellets by the method of extraction used. When fecal samples was used, a large quantity of total RNA serves as carrier of BCoV RNA, demonstrating a high analytical sensitivity and lack of possible substances inhibiting the PCR. The final semi-nested RT-PCR protocol was applied to 25 fecal samples from adult cows, previously tested by a nested RT-PCR RdRp used as a reference test, resulting in 20 and 17 positives for the first and second tests, respectively, and a substantial agreement was found by kappa statistics (0.694). The high sensitivity and specificity of the new proposed method and the fact that primers were designed based on current BCoV sequences give basis to a more accurate diagnosis of BCoV-caused diseases, as well as to further insights on protocols for the detection of other Coronavirus representatives of both Animal and Public Health importance.

Evaluation of a protocol for rapid diagnosis of enterovirus associated with acute flaccid paralysis cases

Background The virological surveillance of acute flaccid paralysis (AFP) is a critical component of the initiative of the World Health Organization (WHO) to eradicate poliomyelitis worldwide. Furthermore rapid methods are needed either to detect or rule out the presence of polioviruses during the late stages of eradication, especially in polio-free areas. Objectives The aim of this study was to evaluate a fast protocol combining one passage (5 days) in cell culture followed by RT-PCR and molecular typing in order to detect and type poliovirus (PV) and other enteroviruses associated with AFP cases. Study design A total of 216 fecal suspensions from AFP suspected cases were tested by using this approach and compared with the WHO gold standard. Results Using the WHO protocol enterovirus was detected in 12 out of the 216 AFP samples (5.55%) while with the proposed protocol enterovirus was detected in 15 out of the 216 AFP samples (6.94%). The additional positive samples detected by the proposed method were classified as non-polio enteroviruses (NPEV). Conclusions The proposed protocol showed higher sensitivity than the WHO gold standard, reducing the entire process of identification and typing of the isolates from the typically 14–21 days to only ∼6–8 days.

Persistence of Human Noroviruses on Food Preparation Surfaces and Human Hands

The human noroviruses (NoV) are the major cause of acute non-bacterial gastroenteritis and are commonly transmitted by foodborne routes. Epidemiological evidence from propagated outbreaks, as well as environmental sampling, suggest that these viruses are environmentally stable. The purpose of this study was to examine the persistence of representative human NoV on the fingertips of volunteers and on commonly used food preparation surfaces. Human fingerpads and surfaces (stainless steel, Formica®, and ceramic) were inoculated with 20% fecal suspensions of Norwalk virus (NV) or Snow Mountain virus (SMV). The virus inocula were recovered by elution at serial time points ranging from 0 to 120 min post-inoculation (for fingerpads) and after up to 42 days (for surfaces). The quantity of detectable viral RNA, expressed as genome equivalent particles (GEP) was evaluated using quantitative real-time RT-PCR (RT-qPCR). The amount of NV RNA on the surface decreased gradually over time, with an average reduction ranging from 1.5 to 2.9 log10 GEP after 21–28 days storage under ambient conditions. SMV showed greater environmental persistence, with a 0.4–1.2 log10 GEP reduction on all three surfaces after 42 days of ambient storage. On fingerpads, the amount of human NoV RNA declined slightly (<0.25 log10) after 15 min and remained relatively unchanged thereafter (through 120 min). These results support the epidemiological evidence that food preparation surfaces and human hands can act as vehicles for human NoV transmission long after the initial contamination event has occurred.

Influence of Coptis Chinensis on pharmacokinetics of flavonoids after oral administration of Radix Scutellariae in rats

Radix Scutellariae (RS) and Coptis Chinensis (CC) are the most popular components in traditional Chinese medicine prescriptions. Flavonoids are the main effective ingredients in RS and berberine is the main effective ingredient in CC. The aim of this study was to determine the influence of CC on the pharmacokinetics of flavonoids following the administration of RS in rats and to investigate the effects of CC on the pharmacokinetic mechanism. Rats were administered RS or RS+CC by intragastric gavage (i.g.). Plasma concentrations of baicalin (baicalein 7-glucuronide) and wogonoside (wogonin 7-glucuronide) were measured by HPLC. Pharmacokinetic parameters were calculated from the plasma concentration time curve. The effect of CC on the metabolism of flavonoids in RS by rat intestinal flora was studied using rat fecal suspensions after ig administration of CC. The effect of berberine on the bi-directional transport of baicalein and baicalin was studied in Caco-2 monolayer models. The effect of CC on the metabolism of baicalein and baicalin by the liver was studied using rat liver microsomes after i.g. administration of CC. Compared to RS alone, RS+CC decreased AUC and C(max) of baicalin and wogonoside. CC decreased the hydrolyzation of baicalin and wogonoside by intestinal flora. Berberine decreased the amount of baicalein transported to the basolateral side and the amount of baicalin which appeared at the basolateral side. The results showed that CC may decrease the bioavailability of baicalin and wogonoside in RS and the mechanism was related to CC decreasing the transport of flavonoid aglycones from the mucosa side to the serosal side and the hydrolyzation of flavonoids by inhibiting intestinal flora.

Comparison of Sheep Ruminal Fluid with Sheep and Horse Faeces as Inoculum for In Vitro Gas Production Measurements

Can, A., Hummel, J., Mobashar, M., Boeser, U. and Südekum, K-H. 2009. Comparison of sheep ruminal fluid with sheep and horse faeces as inoculum for in vitro gas production measurements. J. Appl. Anim. Res., 35: 143–148. A total of 22 ruminant feedstuffs, 14 forages and 8 concentrates, originating from Turkey were used to determine kinetics of in vitro gas production using the Hohenheim gas test (HGT) protocol with multiple readings. Three different inocula sources were ruminal fluid (RF) from fistulated sheep, horse faecal suspension (HFS) and sheep faecal suspension (SFS). Maximum rate of gas production was highest with SFS 2.21 ml/h. The best regression equation for predicting 24 h rumen fluid gas production (ml) was obtained with HFS at 48 h [y = 0.92 ±0.03 × + 9.90 ±1.17; residual standard deviation, 4.43; coefficient of determination, 0.86]. However, more research is required to standardize and optimize the HFS method.

New Saffold Cardiovirus in Children, China

New Saffold Cardiovirus in Children, China

The Bioavailability of Apigenin-7-Glucoside Is Influenced by Human Intestinal Microbiota in Rats

We investigated the impact of human intestinal microbiota on bioavailability of the flavone apigenin-7-glucoside (A7G) by comparing germ-free and human microbiota-associated (HMA) rats. First, the ability of the human intestinal microbiota to convert A7G was proven in vitro by incubating A7G with fecal suspensions. Apigenin, naringenin, and 3-(4-hydroxyphenyl)propionic acid were formed as main metabolites. After application of A7G to germ-free rats, apigenin, luteolin, and their conjugates were detected in urine and feces. In HMA rats, naringenin, eriodictyol, phloretin, 3-(3,4-dihydroxyphenyl)propionic acid, 3-(4-hydroxyphenyl)propionic acid, 3-(3-hydroxyphenyl)propionic acid, and 4-hydroxycinnamic acid in their free and conjugated forms were additionally formed. In whole-blood samples from germ-free and HMA rats, only apigenin conjugates and phloretin, respectively, were detected. The total excretion of A7G and its metabolites within 48 h was similarly low in both germ-free and HMA rats, with 11 and 13% of the A7G dose, respectively. In germ-free rats, A7G metabolites dominated by apigenin and its conjugates were mainly excreted with feces. In contrast, the compounds in HMA rats were predominantly recovered from urine, 3-(4-hydroxyphenyl)propionic acid being the main metabolite. The ability of selected gut bacteria and the host intestinal mucosa to deglycosylate A7G was tested using cell extracts. Apigenin was formed by cytosolic extracts of Eubacterium ramulus and Bacteroides distasonis and by the microsomal fraction of the small intestinal mucosa of rats. Overall, human intestinal microbiota largely contributed to A7G metabolism, indicating its influence on the bioactivity of flavones.

Conversion of Dehydrodiferulic Acids by Human Intestinal Microbiota

Plant cell wall associated dehydrodiferulic acids (DFA) are abundant components of cereal insoluble dietary fibers ingested by humans. The ability of human intestinal microbiota to convert DFA was studied in vitro by incubating 8-O-4- and 5-5-coupled DFA with fecal suspensions. 8-O-4-DFA was completely degraded by the intestinal microbiota of the majority of donors, yielding homovanillic acid, 3-(3,4-dihydroxyphenyl)propionic acid, and 3,4-dihydroxyphenylacetic acid as the main metabolites. The transient formation of ferulic acid and presumably 3-(3-hydroxy-4-methoxyphenyl)pyruvic acid suggests an initial cleavage of the ether bond. In contrast to 8-O-4-DFA, the 5-5-coupled DFA was not cleaved into monomers by any of the fecal suspensions. Only the side chains were hydrogenated and the methoxy groups were demethylated. The cleavage of DFA by human intestinal microbiota, which depended on their coupling type, may affect both the bioavailability of DFA and the degradability of DFA-coupled fiber in the gut.

Simian hepatitis A virus derived from a captive rhesus monkey in India is similar to the strain isolated from wild African green monkeys in Kenya

A simian hepatitis A virus (HAV) was identified retrospectively in a faecal sample from a rhesus monkey in India, inoculated in 1995 with a faecal suspension from a suspected patient of non-A to E hepatitis. The monkey was in captivity for 2 years in one of the experimental primate facilities in western India before being moved to the National Institute of Virology, Pune for experimentation. Phylogenetic analysis based on a partial sequence of the 5' noncoding region placed this virus in genotype V, the only other member being the AGM-27 strain recovered in 1986 from African green monkeys in Kenya. The source of infection of the monkey remains unclear. The full genome was amplified in nine fragments and sequenced. The genome of the Indian simian HAV (IND-SHAV) is 7425 nucleotides long including the poly-A tail of 14 nucleotides at the 3' end. At the nucleotide and amino acid levels, IND-SHAV was 99.8 and 100% identical with AGM27, respectively.