Supplementation with psyllium seed husk reduces myocardial damage in a rat model of ischemia/reperfusion

BACKGROUND/OBJECTIVES: Myocardial infarction (MI) is caused by extensive myocardial damage attributed to the occlusion of coronary arteries. Our previous study in a rat model of ischemia/reperfusion (I/R) demonstrated that administration of arabinoxylan (AX), comprising arabinose and xylose, protects against myocardial injury. In this study, we undertook to investigate whether psyllium seed husk (PSH), a safe dietary fiber containing a high level of AX (> 50%), also imparts protection against myocardial injury in the same rat model. MATERIALS/METHODS: Rats were fed diets supplemented with PSH (1, 10, or 100 mg/kg/d) for 3 d. The rats were then subjected to 30 min ischemia through ligation of the left anterior descending coronary artery, followed by 3 h reperfusion through release of the ligation. The hearts were harvested and cut into four slices. To assess infarct size (IS), an index representing heart damage, the slices were stained with 2,3,5-triphenyltetrazolium chloride (TTC). To elucidate underlying mechanisms, Western blotting was performed for the slices. RESULTS: Supplementation with 10 or 100 mg/kg/d of PSH significantly reduces the IS. PSH supplementation (100 mg/kg/d) tends to reduce caspase-3 generation and increase BCL-2/BAX ratio. PSH supplementation also upregulates the expression of nuclear factor erythroid 2-related factor 2 (NRF2), and its target genes including antioxidant enzymes such as glutathione S-transferase mu 2 (GSTM2) and superoxide dismutase 2 (SOD2). PSH supplementation upregulates some sirtuins (NAD+-dependent deacetylases) including SIRT5 (a mitochondrial sirtuin) and SIRT6 and SIRT7 (nuclear sirtuins). Finally, PSH supplementation upregulates the expression of protein kinase A (PKA), and increases phosphorylated cAMP response element-binding protein (CREB) (pCREB), a target protein of PKA. CONCLUSIONS: The results from this study indicate that PSH consumption reduces myocardial I/R injury in rats by inhibiting the apoptotic cascades through modulation of gene expression of several genes located upstream of apoptosis. Therefore, we believe that PSH can be developed as a functional food that would be beneficial in the prevention of MI.

Pentoses Used in Cultures of Synechococcus nidulans and Spirulina paracas: Evaluation of Effects in Growth and in Content of Proteins and Carbohydrates

Abstract The biological assimilation of the sugars present in lignocellulosic residues has gained prominence since these residues are the most abundant and economic residues in nature. Thus, the objective of this work was to determine whether the use of D-xylose and L-arabinose as sources of carbon in Synechococcus nidulans and Spirulina paracas cultures affects the growth and production of proteins and carbohydrates. Kinetic growth parameters, pentose consumption, protein content and carbohydrates were evaluated. Synechococcus nidulans and Spirulina paracas consumed all concentrations of pentose used. The highest cellular concentration (1.37 g.L-1) and the highest protein productivity (54 mg.L-1.d-1) were obtained for Spirulina paracas, which was submitted to the addition of 38.33 mg.L-1 D-xylose and 1.79 mg.L-1 L-arabinose. The use of pentose promoted the accumulation of proteins for the studied microalgae. This is one of the first works to report protein bioaccumulation as a result of pentose addition.

Production optimization, purification, expression, and characterization of a novel α-L-arabinofuranosidase from Paenibacillus polymyxa

Background: α-L-Arabinofuranosidase (EC 3.2.1.55) catalyzes the hydrolysis of terminal α-L-1,2-, -1,3-, and -1,5- arabinofuranosyl residues in arabinose-containing polymers, and hence, it plays an important role in hemicellulose degradation. Herein, the bacterium Paenibacillus polymyxa, which secretes arabinofuranosidase with high activity, was selected for enzyme production, purification, and characterization. Results: Medium components and cultural conditions were optimized by the response surface method using shake flask cultures. Arabinofuranosidase production reached 25.2 U/mL under optimized conditions, which were pH 7.5, 28°C, and a basic medium supplemented with 1.5 g/L mannitol and 3.5 g/L soymeal. Furthermore, the arabinofuranosidase secreted by P. polymyxa, named as PpAFase-1, was partially purified from the supernatant using a DEAE Sepharose Fast Flow column and a hydroxyapatite column. The approximate molecular mass of the purified PpAFase-1 was determined as 56.8 kDa by SDS-PAGE. Protein identification by mass spectrometry analysis showed that the deduced amino acid sequence had significant similarity to the glycosyl hydrolase family 51. The deduced gene of 1515 bp was cloned and expressed in Escherichia coli BL21 (DE3) cells. Purified recombinant PpAFase-1 was active toward p-nitrophenyl-α-L-arabinofuranoside (pNPAraf). The Km and kcat values toward pNPAraf were 0.81 mM and 53.2 s−1 , respectively. When wheat arabinoxylan and oat spelt xylan were used as substrates, PpAFase-1 showed poor efficiency. However, a synergistic effect was observed when PpAFase-1 was combined with xylanase from Thermomyces lanuginosus. Conclusion: A novel GH51 enzyme PpAFase-1 was cloned from the genome of P. polymyxa and expressed in E. coli. This enzyme may be suitable for hemicellulose degradation on an industrial scale.

Protection of the brain through supplementation with larch arabinogalactan in a rat model of vascular dementia

BACKGROUND/OBJECTIVES: Vascular dementia (VaD) caused by reduced blood supply to the brain manifests as white matter lesions accompanying demyelination and glial activation. We previously showed that arabinoxylan consisting of arabinose and xylose, and arabinose itself attenuated white matter injury in a rat model of VaD. Here, we investigated whether larch arabinogalactan (LAG) consisting of arabinose and galactose could also reduce white matter injury. MATERIALS/METHODS: We used a rat model of bilateral common carotid artery occlusion (BCCAO), in which the bilateral common carotid arteries were exposed and ligated permanently with silk sutures. The rats were fed a modified AIN-93G diet supplemented with LAG (100 mg/kg/day) for 5 days before and 4 weeks after being subjected to BCCAO. Four weeks after BCCAO, the pupillary light reflex (PLR) was measured to assess functional consequences of injury in the corpus callosum (cc). Additionally, Luxol fast blue staining and immunohistochemical staining were conducted to assess white matter injury, and astrocytic and microglial activation, respectively. RESULTS: We showed that white matter injury in the the cc and optic tract (opt) was attenuated in rats fed diet supplemented with LAG. Functional consequences of injury reduction in the opt manifested as improved PLR. Overall, these findings indicate that LAG intake protects against white matter injury through inhibition of glial activation. CONCLUSIONS: The results of this study support our hypothesis that cell wall polysaccharides consisting of arabinose are effective at protecting white matter injury, regardless of their origin. Moreover, LAG has the potential for development as a functional food to prevent vascular dementia.

Ethanol Production from Various Sugars and Cellulosic Biomass by White Rot Fungus Lenzites betulinus

Lenzites betulinus, known as gilled polypore belongs to Basidiomycota was isolated from fruiting body on broadleaf dead trees. It was found that the mycelia of white rot fungus Lenzites betulinus IUM 5468 produced ethanol from various sugars, including glucose, mannose, galactose, and cellobiose with a yield of 0.38, 0.26, 0.07, and 0.26 g of ethanol per gram of sugar consumed, respectively. This fungus relatively exhibited a good ethanol production from xylose at 0.26 g of ethanol per gram of sugar consumed. However, the ethanol conversion rate of arabinose was relatively low (at 0.07 g of ethanol per gram sugar). L. betulinus was capable of producing ethanol directly from rice straw and corn stalks at 0.22 g and 0.16 g of ethanol per gram of substrates, respectively, when this fungus was cultured in a basal medium containing 20 g/L rice straw or corn stalks. These results indicate that L. betulinus can produce ethanol efficiently from glucose, mannose, and cellobiose and produce ethanol very poorly from galactose and arabinose. Therefore, it is suggested that this fungus can ferment ethanol from various sugars and hydrolyze cellulosic materials to sugars and convert them to ethanol simultaneously.

Biohydrogen production by Thermoanaerobacterium thermosaccharolyticum KKU-ED1: Culture conditions optimization using mixed xylose/arabinose as substrate

Background: Biological hydrogen production by microorganisms can be divided into two main categories i.e. photosynthetic organisms that produce hydrogen using light as energy source and anaerobic bacteria that produce hydrogen via dark fermentation. Dark fermentative hydrogen production by anaerobic bacteria has the advantages of a higher HPR without illumination and of the capability to convert various kinds of substrate. Results: Thermophilic hydrogen producer was isolated from elephant dung and identified as Thermoanaerobacterium thermosaccharolyticum KKU-ED1 by 16S rRNA gene analysis, which was further used to produce hydrogen from mixed pentose sugar i.e., xylose/arabinose. The optimum conditions for hydrogen production from mixed xylose/arabinose by KKU-ED1 were a 1:1 xylose/arabinose mixture at the total concentration of 5 g/L, initial pH of 6.5 and temperature of 55ºC. Under the optimum conditions, hydrogen from sugar derived from acid-hydrolyzed sugarcane bagasse at a reducing sugar concentration were achieved. Soluble metabolite product (SMP) was predominantly acetic acid indicating the acetate-type fermentation. Conclusions: The strain KKU-ED1 appeared to be a suitable candidate for thermophilic fermentative hydrogen production from hemicellulosic fraction of lignocellulosic materials due to its ability to use various types of carbon sources.

Applicability of a natural swelling matrix as the propellant of osmotic pump tablets / 药学学报

The purpose of this study is to investigate the applicability of a natural swelling matrix derived from boat-fruited sterculia seed (SMS) as the propellant of osmotic pump tablets. The sugar components, static swelling, water uptake and viscosity of SMS were determined and compared with that of polythylene oxide (WSR-N10 and WSR-303). Both ribavirin and glipizide were used as water-soluble and water-insoluble model drugs. Then, the monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide were prepared using SMS as the osmotically active substance and propellant. SMS was mainly composed of rhamnose, arabinose, xylose and galactose and exhibited relatively high swelling ability. The area of the disintegrated matrix tablet was 20.1 times as that at initial after swelling for 600 s. SMS swelled rapidly and was fully swelled (0.5%) in aqueous solution with relative low viscosity (3.66 +/- 0.03) mPa x s at 25 degrees C. The monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide using SMS as propellant exhibited typical drug release features of osmotic pumps. In conclusion, the swelling matrix derived from boat-fruited sterculia seed, with low viscosity and high swelling, is a potential propellant in the application of osmotic pump tablets.

Development of a new HPLC technique for analyzing monosaccharide composition and its application in the quality control of Silybum marianum polysaccharide / 药学学报

A new HPLC-UV technique for the separation and analysis of 10 monosaccharides achieved within 13.5 min using 1-phenyl-3-methyl-5-pyrazolone (PMP) as the labelling molecule of the reductive monosaccharides has been established by combining common high performance liquid chromatography-UV and C18 column. The established technique was applied to the quantification of the monosaccharide components in extract of Silybum marianum. The results showed that the tested 10 monosaccharides as PMP derivatives were baseline separated under the HPLC conditions proposed. It was confirmed that Silybum marianum extract was composed of mannose, rhamnose, glucuronic acid, galacturonic acid, glucose, xylose, galactose and arabinose with the molar ratio of 0.66:0.84:0.58:1.0:1.6:0.69:2.7:4.8. Quantitative recoveries of the compositional monosaccharides separated from the extract were in the range of 92.4%-104.0%, and the RSD values fell within 0.68%-3.81%. The results demonstrated that the proposed HPLC method was simple, rapid, convenient, and precise, and it was applicable to the analysis of the compositional monosaccharides of Silybum marianum extract.

Analysis of monosaccharides in Radix Rehmanniae by GC / 中国中药杂志

<p><b>OBJECTIVE</b>To isolate and purify the polysaccharides from Radix Rehmanniae and analysis the monosaccharides composition.</p><p><b>METHOD</b>The polysaccharides were extracted with hot water and precipitated by alcohol. Proteins in the precipitates were removed by TCA method. The products were further purified with column chromatography on Superdex 200 and Sephadex G100. The SRP I and SRP II were identified as homogeneous polysaccharide by HPLC, respectively, and then analyzed by GC after being hydrolysised.</p><p><b>RESULT</b>Two homogeneous polysaccharides (SRP I and SRP II) were obtained from Radix Rehmanniae.</p><p><b>CONCLUSION</b>SRP I contained rhamnose, arabinose, glucose and galactose in the percentage of 6.11%, 66.46%, 3.93% and 21.50%. SRP I was composed of rhamnose, fucose, mannose, galactose and fructose in the percentage of 21.82%, 24.47%, 10.48%, 29.94% and 13.29%.</p>

Composition and Partial Structure Characterization of Tremella Polysaccharides

Heteropolysaccharides isolated from liquid cultures of nine Tremella species contained 0.3 to 1.2% protein, 2.7 to 5% ash, 0.9 to 3.4% acetyl groups, 76.5 to 84.2% carbohydrates and trace amounts of starch. The polysaccharides in aqueous solution were slightly acidic (pH 5.1 to 5.6). They consisted of the following monomeric sugars: fucose, ribose, xylose, arabinose, mannose, galactose, glucose and glucuronic acid. The backbones of the polysaccharide structures consisted of alpha-(1-->3)-links while the side chains were beta-linked.

Development of Yvrk Gene-Recombinant E. coli Degrading Oxalate

PURPOSE: Recently, the whole DNA sequence of Bacillus subtilis (B. subtilis) was identified, revealing the existence of the YvrK gene encoding a 43 kD oxalate decarboxylase (OXDC), which degrades oxalate by a simple pathway. The objective of this study was to develop recombinant Escherichia coli (E. coli) expressing the Yvrk gene from B. subtilis. MATERIALS AND METHODS: After the extraction of total DNA from B. subtilis, the YvrK gene was cloned by polymerase chain reaction. The cloned DNA encoding OXDC was inserted into the pBAD/gIII-A vector, downstream of the L-arabinose promotor. The plasmid vector was transformed into TOP 10 E. coli, and the transformants were selected with ampicillin. The recombinant E. coli, named pBy, was then analyzed by DNA sequencing and Western blot. To evaluate the oxalate-degrading function of pBy, pBy was cultured in LB broth containing oxalate, and then the amount of oxalate in the medium was assessed. The oxalate-degrading activity of homogenates of pBy was evaluated. RESULTS: DNA sequencing showed the successful transformation of the YvrK gene into TOP 10 E. coli. Western blot analyses showed that pBy expressed OXDC. pBy removed oxalate during the overnight culture in oxalate-containing LB broth, and the homogenate of pBy degraded 90% of oxalate under acidic conditions. CONCLUSIONS: A recombinant E. coli expressing the YvrK gene was successfully produced. The bacteria showed potent oxalate-degrading activity. The results of this study will provide a solution to the treatment of calcium oxalate stones and hyperoxaluria, for which there are few medical treatment modalities.

Cloning, optimization of expression condition, purification and immunological characterization of hydrophilic section of HCV core Ag, expressed in E-coli by T7-ara BAD promoter

The capsid or core Ag of Hepatitis C virus is a multifunctional protein which has the principal pathogenesis and diagnostic role in HCV related infections and most of these properties are attributed to the hydrophilic section [amino acids 2-122] of this protein. For different research and diagnostic applications, high amounts of this protein in pure and original form are required. So, the aim of this study was to clone the gene, optimize the expression condition, purify it in the original form, and immunologically characterize hydrophilic section of HCV Core Ag, expressed by T7-araBAD promoter system in E.coli. The PCR amplified region corresponding to 2-122 section of this Ag from genotype lb was cloned in pIVEX 2.3, a T7 promoter derived vector. The proper construct after digestional analysis and sequencing confirmations was transformed into BL2 1 -Al E. coli, and protein expression under control of araBAD promoter by addition of 0.2% Arabinose was induced. After optimization of expression condition, purification of protein by NI-NTA agarose gel chromatography in native condition by immidazole yielded about 3.5mg/L of HCV core Ag. Immunological studies by western blotting through application of core specific mAbs and results of ELISA tests indicated that the protein is with desired immunological properties. AraBAD promoter can be perfectly utilized to produce the hydrophilic section of HCV core in high yields, and purification through NI-NTA in native condition may provide the antigen for different research and diagnostic applications

Isolation and structure elucidation of glycosides in n-butanol extracts from rhizome of Periploca calophylla / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from n-butanol extracts of Periploca calophylla.</p><p><b>METHOD</b>The constituents were isolated and purified by chromatographic technology and their structures were elucidated on the basis of physicochemical property and spectroscopic methods.</p><p><b>RESULT</b>Eight glycosides were isolated and identified as periplocin (I), kaempferol 3-alpha-D-arabinoside (II), kaempferol 3-O-beta-D-glucoside (III), 3',4',5,7-tetrahydroxyflavanone-2(S)-3'-O-beta-D-glucopyranoside (IV), (+)-syringaresinol-4'-O-beta-D-monoglucoside (V), 1-sinapoylglucoside (VI), erigeside C (VII), 2,6-dimethoxy-4-hydroxyphenol 1-O-beta-D-glucoside (VIII).</p><p><b>CONCLUSION</b>All the compounds were isolated for the first time from this plant.</p>

Identification of Enterococcus Species Using a Microplate / 임상검사와정도관리

BACKGROUND: The aim of the study was to develop an accurate, convenient, and easy microplate system for the identification of enterococcal species from clinical specimens. METHODS: The microplate identification method was composed of twelve biochemical tests and identification programs. The tests comprised in microplate were initially screened by a two-tube method, NaCl-esculin hydrolysis and pyrrolidonyl-beta-naphthylamide test; arginine dihydrolase, acid production from mannitol, sorbitol, sucrose, arabinose, raffinose, methyl-alpha-D-glucopyranoside, and ribose in the microplate; and pigment production and hemolytic pattern in blood agar plate. The performance of the microplate for identifying enterococci to the species level was evaluated in comparison with conventional reference tests and commercial kits. RESULTS: Among the 111 clinical isolates of Enterococcus species, the microplate system correctly identified 100% to genus level, and 91.0% to species level. All of E. casseliflavus, E. durans, and E. hirae were correctly identified by the microplate. The diagnostic sensitivity and specificity for identification of Enterococcus species were as follows: 100% and 96.7% in E. faecium, 93.5% and 100% in E. faecalis, 100% and 97.2% in E. raffinosus, and 33.3% and 98.1% in both E. avium and E. gallinarum. CONCLUSIONS: It is concluded that the microplate method offers a simple, cost-effective, rapid, and accurate identification system for the identification of most clinical isolates of Enterococcus species.

Chemical constituents of Geranium eristemon / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents of Geranium eristemon.</p><p><b>METHOD</b>Chromatography and spectral analysis were used to isolate the constituents and elucidate their structures.</p><p><b>RESULT</b>Five compounds were isolated from acetone extract of the whole grass of G. eristemon, and identified as beta-sitosterol, protocatechuic acid, myricetin, kaempferol-7-O-alpha-L-arabifuranoside and kaempferol-3-O-alpha-L-arabifuranoside.</p><p><b>CONCLUSION</b>kaempferol-7-O-alpha-L-arabifuranoside and kaempferol-3-O-alpha-L-arabifuranoside were isolated from G. genus for the first time.</p>

Structural features of a neutral heteropolysaccharide CPB-4 from Cynanchum paniculatum / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical features of CPB-4, a heteropolysaccharide obtained from Cynanchum paniculatum.</p><p><b>METHOD</b>Sugar composition analysis, methylation analysis, partial hydrolysis and carbon-13 nuclear magnetic resonance were used to determine the sugar composition, linkages, main chain, branch chains and branching points.</p><p><b>RESULT</b>CPB-4 is composed of L-arabinose, L-xylose, L-rhamnose and D-galactose in closely molar ratios of 0.8:0.2:0.2:1.0. Its main chain is comprised of 1, 5 linked galactose and side chains are comprised of terminal xylose, terminal arabinose, oligosaccharide of arabinose and oligosaccharide of arabinose, rhamnose and galactose. The branching points are located at C-6 and C-2 of galactose.</p><p><b>CONCLUSION</b>CPB-4 is a new heteropolysaccharide from C. paniculatum.</p>

Chemical studies on SC3, a polysaccharide from Salvia chinensis / 药学学报

<p><b>AIM</b>To study the chemical structure of SC3, an acidic polysaccharide from Salvia chinesis.</p><p><b>METHODS</b>Based on chemical (including sugar composition analysis, methylation analysis, uronic acid reduction and partial acid hydrolysis) and spectral analysis (IR, NMR, ESI-MS), the structural characterization of SC3 was investigated.</p><p><b>RESULTS</b>SC3 was composed Rha, Ara, Gal and GalA, with its mean molecular weight of 7.7 x 10(4). By means of methylation analysis, partial acid hydrolysis, NMR and ESI-MS spectrum, the linkages and sequence information of SC3 were obtained.</p><p><b>CONCLUSION</b>SC3 is an complicated acidic polysaccharide, obtained for the first time from the plant.</p>

Isolation and identification of Vibrio vulnificus and Vibrio parahaemolyticus from coast of Pusan and Daechon

This study was focused on the isolation of pathogenic Vibrio species, V. vulnificus and V. parahaemolyticus from marine environment from May to July of 1999. Isolation sites were coast near by Pusan and Daechon. The results obtained were as follows: 1. Seventy strains of V. parahaemolyticus and 19 strains of V. vulnificus were isolated from a total of 120 specimens. 2. Nineteen strains of V. vulnificus did not fermented arabinose and salicin but fermented lactose and cellobiose. All of V. parahaemolyticus isolates did not fermented lactose and cellobiose. 47 strains of V. parahaemolyticus fermented arabinose but 53 strains did not fermented salicin. 3. V. vulnificus and V. parahaemolyticus isolates showed three different API index numbers with 5046105 and 4346107 dominant. 4. V. vulnificus did not grow on 0% and 8% NaCl containing medium. V. parahaemolyticus grew on 8% NaCl containing medium. 5. V. vulnificus isolates and V. parahaemolyticus revealed different outer membrane protein p rofiles on SDS-PAGE.

Epidemiology of arabinose assimilation in burkholderia pseudomallei isolated from patients and soil in Thailand.

Burkholderia pseudomallei is an environmental saprophyte that has been isolated widely from soil in Southeast Asia and the relationship between environmental contamination and clinical melioidosis has been established. It has been shown that the arabinose assimilation property of B. pseudonrallei is probably one of the determinants indicating virulence of this organism. Therefore, the distribution of arabinose assimilation biotypes of B. pseudomallei collected from four geographic regions of Thailand was studied in order to determine an association between arabinose assimilation of B. pseudomallei and the uneven distribution of melioidosis found among these four areas. A total of 830 isolates of B. pseudomallei (412 patient isolates and 418 soil isolates) collected from the patients and soil in four regions of Thailand in 1997 were tested for an ability to grow on a minimal agar medium supplemented with L-arabinose. All patient isolates except one could not utilise arabinose (Ara-). For 418 soil isolates, 232 (55.5%) isolates were identified as Ara type. They comprised 180 (62.5%), 36 (46.8%), 6 (35.3%) and 10 (27.8%) isolates derived from northeastern, southern, northern and central regions respectively. The ratios of Ara- to Ara, were 1.7, 0.9. 0.5 and 0.4 among isolates collected from northeastern, southern, northern and central regions respectively. The prevalence of Ara- in soil isolates in northeast is significantly higher than those in other regions. This observation suggests that in addition to the presence of B. pseudomallei in soil which is one of the factors contributing to a burden of melioidosis in northeastern Thailand, the distribution of more virulent biotype (Ara-) soil isolates is a factor contributing to a high prevalence of melioidosis in northeastern Thailand as well.

Development of Simple Identification Method of Enterococci / 대한임상병리학회지

BACKGROUND: The selection of identification (ID) system of enterococci depends mainly on the accuracy of ID system, cost of operation and convenience of testing. Commercial ID kits are easy to use but too expensive. The aim of the study was to develop a simple system for the identification of species of enterococci which are frequently isolated from clinical specimens. METHODS: Eight conventional biochemical tests selected for the simple ID method were hemolysis pattern, NaCl-esculin hydrolysis, tellurite tolerance, arginine dihydrolase, acid from arabinose, raffinose and methyl-alpha-D-glucopyranoside, and pigment production. Ninety one consecutive strains of enterococci from clinical specimens isolated during the period of April 1988 were tested by the simple ID method, and API rapid ID 32 STREP. RESULTS: Simple ID method with 15 ID codes was established to identify 13 species of enterococci. Among the 91 isolates tested, 88 (96.7%) were identified to the species level of enterococci by simple ID method. CONCLUSIONS: The simplified conventional ID method is simple, reliable and economical. Further modification may improve the accuracy of the enterococcal species identification system.