Screening and Enzymatic Activity Analysis of Dominant Fermentive Bacteria of Sojae Semen Praeparatum from Different Production Places / 中国药房

OBJECTIVE:To provide reference for the standardized production of Sojae semen praeparatum (SSP). METH-ODS:SSP samples from Heilongjiang,Hebei,Gansu,Shandong,Anhui and Yunnan were respectively collected. The fermentive bacteria were cultured with the selective medium contained artemisiae annuae herba and mori folium. Foline-phenol method,fibrous protein plate method and p-nitrophenol-β-D-glucoside colorimetric method were respectively conducted to determine the activities of protease,plasmin and β-glucosidase of the strains to screen dominant fermentive bacteria. RESULTS:Totally 14 wild strains were separated from SSP samples from 6 production places, including 3 strains of bacteria and 11 strains of molds. 1 strain of rod-shaped bacteria and 1 strain of Mucor sp. were separated from SSP from Heilongjiang;2 strains of Mucor sp. and 1 strain of rod-shaped bacteria were separated from SSP from Hebei;1 strain of Mucor sp.,1 strain of Penicillium sp.,1 strain of Streptococ-cus sp. and 1 strain of Aspergillus sp. were separated from SSP from Gansu;2 strains of Mucor sp. were separated from SSP from Shandong;1 strain of Mucor sp. and 1 strain of Aspergillus sp. were separated from SSP from Anhui;and only 1 strain of Mucor sp. was separated from SSP from Yunnan. According to the strains category and enzyme activities,No.1 bacillus,No.9 Aspergillus sp.,No.11 and No.14 Mucor sp. were preliminary authenticated as dominant fermentation microorganism,total enzyme activities of the 4 strains were 22.77,25.49,41.32,39.13 U/g respectively. CONCLUSIONS:The fermentive bacteria of SSP from different pro-duction places were different,and the dominant one can be screened preliminary through enzyme activity analysis.

Preliminary screening for in vitro anti-enteritic properties of a traditional herb Dillenia pentagyna Roxb. fruit extracts

OBJECTIVE@#To explore anti-enteric properties of Dillenia pentagyna Roxb. (D. pentagyna) fruit extract fractions of different polarities by comparative antimicrobial activity against municipal sewage microflora and to assess its urease inhibition potential.@*METHODS@#Different polar fractions of D. pentagyna fruit extracts were studied by antimicrobial susceptibility test with several adjustments in this resource limited setup. Tests were done against municipal sewage microbes at various bacterial load (initially with 1.0 McFarland followed by 1.5 McFarland) using basal (nutrient agar) and selective medium (MacConkey's agar with and without supplementation of 5% NaCl). All assays statistically scaled with a gradient for uniformity and comparison with ciprofloxacin standard. Fraction with highest activity was studied for urease inhibition potential by kinetic method.@*RESULTS@#DP-47 separated by 30% ethyl acetate (EtOAc) in CHCl3 from CHCl3 extract had slightly higher antimicrobial potency (y=0.758x+7.571) as dissolved in methanol than in dimethylsulfoxide (y=0.300x+6.000). EtOAc extract fractioned by 10% methanol in EtOAc (DP-43) was more potent antimicrobial (y=1.428x+8.392) and soluble in water. Butanol extract fractioned by water (DP-50) showed highest antimicrobial potency (y=3.384x+2.000) than both DP-47 and DP-43 in disk diffusion assays. In higher microbial load DP-50 was potent antimicrobial (y=1.538x+3.000) and completely inhibited any vegetative growth at lower load of 0.5 McFarland. In selective environment DP-50 was effective (y=1.076x+7.500 in MacConkey's, y=1.307x+6.000 in 5% NaCl supplemented). It was evident that enteric pathogens may not easily develop resistance against DP-50 and at high concentration it inhibited urease activity by 94.87%. The standard inhibition by thiourea (1%) is 33.914%.@*CONCLUSIONS@#Highly polar fraction of D. pentagyna Roxb. fruit extract DP-50 have potential antimicrobial activity against sewage microflora in basal and selective culture indicating its potential against non-fastidious, coliforms and Vibrio pathogens. Urease inhibition indicates efficacy against Helicobactor pylory.

Preliminary screening for in vitro anti-enteritic properties of a traditional herb Dillenia pentagyna Roxb. fruit extracts

Objective: To explore anti-enteric properties of Dillenia pentagyna Roxb. (D. pentagyna) fruit extract fractions of different polarities by comparative antimicrobial activity against municipal sewage microflora and to assess its urease inhibition potential. Methods: Different polar fractions of D. pentagyna fruit extracts were studied by antimicrobial susceptibility test with several adjustments in this resource limited setup. Tests were done against municipal sewage microbes at various bacterial load (initially with 1.0 McFarland followed by 1.5 McFarland) using basal (nutrient agar) and selective medium (MacConkey's agar with and without supplementation of 5% NaCl). All assays statistically scaled with a gradient for uniformity and comparison with ciprofloxacin standard. Fraction with highest activity was studied for urease inhibition potential by kinetic method. Results: DP-47 separated by 30% ethyl acetate (EtOAc) in CHCl

Studies on strains of Trichoderma spp. from high altitude of Garhwal Himalayan region.

Two species of Trichoderma i.e. T. harzianum and T. viride have been isolated from the soil samples collected from the higher altitude (2000-3500 m) of Garhwal Himalayan region in India. The two species were grown in Petri plates on TSM agar media and it was also observed that the optimum temperature and pH for Trichoderma growth was 300 C and 5.5 respectively. When incubated on TSM agar medium at 40 C, the fungus grew normally with heavy induced sporulation within three weeks of incubation. Induction of sporulation on exposure to low temperature appeared to be strategies for survival of these species in extreme cold environment temperature 4 to 50 C. Antifungal activities of the two species of Trichoderma were demonstrated with phytopathogenic fungi in dual cultures. The antifungal metabolites produced by Trichoderma spp., diffusible as well as volatile, caused abnormalities in pathogenic fungi. Plant growth promotion of Trichoderma spp. was also shown through plant analysis in greenhouse.

The Composition of a Selective Media for Trichomonas vaginalis Isolation / 감염

BACKGROUND: Modified Diamond medium (MDM) supplemented with 10% heat-inactivated horse serum, streptomycin, penicillin G, and mycostatin is commonly used for the isolation of Trichomonas vaginalis from vaginal swab. But, judging from our experience, the above usual MDM antibiotic composition was frequently contaminated with facultative anaerobes, and isolation rate of T. vaginalis was no more than 12% in 142 korean woman patients whose chief complaints were foul odored, increased vaginal discharge. This isolation rate is low in comparison with reports of another countries including U.S.A (about 15~30%) and could be attributed to the prevalence of antibiotic resistance in Korea. So, we exploited more selective antibiotic compositions in modified Diamond medium for pure isolation of T. vaginalis. METHODS: We used new self-devised anaerobic pack for sample maintenance and tested several antibacterial and antifungal agent combinations in modified Diamond medium supplemented with 5% human erythrocyte lysate and 5% heat-inactivated human serum in the place of 10% horse serum with the object of increased and pure isolation of T. vaginalis. Several drugs and chemicals were tested to fourteen wild strains isolated in a local clinic, in the hope of finding the agents that have no effect on T. vaginalis growth in high drug concentrations. Anaerobic jar was used for culture of T. vaginalis and cell count performed in the improved Neubauer's haemocytometer. RESULTS: Strains of T. vaginalis grew batter in modified Diamond medium supplemented with 5% human erythrocyte lysate and 5% heat-inactivated human serum (mean 1.166X106, about 5.83 fold) than 10% horse serum (mean 2.0X105 after 48 hours culture), and their growth rate in the former was more rapid than the latter in early grow phase. On the basis of this results, we examined selectivity of modified Diamond media supplemented with several antibacterial and antifungal combinations by a double blind test. Isolation rate in the conventional modified Diamond's medium (combination A; 10% horse serum, streptomycin 1,200 microgram/mL, penicillin G 1,500 unit/mL, mycostatin 37.5 microgram/mL, pH 6.2) was 9/73 (12.3%) while in modified Diamond medium supplemented with 5% human erythrocyte lysate and 5% heat-inactivated human serum, isolation rates in various drug combinations were as follows; Combination B (cefazolin 100 microgram/mL, streptomycin 1,200 microgram/mL, clindamycin 150 microgram/mL, pH 6.5), combination C (bacitracin 14.6 unit/mL, streptomycin 1,200 microgram/mL, clindamycin 150 microgram/mL pH 6.5) and combination D (vancomycin 100 microgram/mL, streptomycin 1,200 microgram/mL, clindamycin 150 microgram/mL pH 6.5) were all 11/73 (15.0%). Combination D allowed the least bacterial growth rate. CONCLUSION: We consider that a new modified Diamond medium supplemented with 5% human erythrocyte lysate, 5% heat-inactivated human serum and combination D might be provide the highest selection for Trichomonas vaginalis pure isolation from vaginal swabs.