Abstract: Objective This study assesses the antitubercular potential of natural products obtained from plants reputed to have medicinal properties and collected from the tropical flora of Puerto Rico. Background The increase in persons infected with Mycobacterium tuberculosis (MTB) the world over and the development of resistance to antibiotics by this microbe and other infectious bacteria has created the need for new drugs to replace those which have lost effectiveness. Method In Phase I of this study, ethanolic leaf extracts of fifty local plants were submitted to preliminary screening to assess their in vitro Mycobacterium smegmatis inhibitory activity using the Bauer-Kirby disk diffusion method. In Phase II, the definitive screening of the six most promising extracts which inhibited M. smegmatis were assayed for their MTB inhibitory activity using the BACTEC 460 susceptibility test method. The brine shrimp bioassay was used as a toxicity bioassay and the mice inoculation test was used to determine mice tolerance to the effect of the daily intraperitoneal inoculations of the plant extracts. Results MTB showed varying degrees of susceptibility to each plant extract. This effect was dependent upon the plant species, dose and time of exposure. Evidence is provided suggesting that: (1) Six crude plant extracts (12%) tested possessed inhibitory capacity at the amount of 500 micrograms per disc; (2) Mammea americana extract yielded the strongest inhibitory effect at 50 micrograms per disc, followed by Marchantia polymorpha, Mangifera indica, Callistemon citrinus, Syzygium jambos and Momordica charantia; (3) the bactericidal inhibitory pattern of MTB growth, exposed to Mammea americana extract, was comparable to streptomycin; and (4) the transitory reduction pattern of MTB growth, produced by Callistemon citrinus, Marchantia polymorpha extracts at 100 micrograms and 250 micrograms, was similar to that of bacteriostatic agents. Conclusion Of 50 plants screened six extracts tested for their anti-MTB activity yielded positive results with varying degrees of inhibition. Mammea americana showed the greatest inhibitory activity suggesting that certain plant species yield valuable anti-Mycobacterium tuberculosis substances. The procedures employed in this study, including the BACTEC 460 modified method, are useful for in vitro screening of plant extracts with potential antitubercular activity.

Abstract: Firouzi, R, Azadbakht, M. and Nabinedjad, A. 1998. Anti-listerial activity of essential oils of some plants. J. Appl. Anim. Res., 14: 75–80. Minimal inhibitory concentration (MIC) of essential oils of plant Thymus daensis, Artemisia absintium, Matricaria chamommilla, Melissa officinalis and Allium sativum (garlic) against Listeria monocytogenes type 4a using tube dilution technique and agar disk diffusion method were studied In tube dilution technique the concentration of each oil was estimated in v/v units. The results showed that the MIC values for T. daensis, A absintium, M. chamommilla, M. officinalis and A. sativum, were 1:6400, 1:1280, 1:1280, 1:2560 and 1:1280, respectively. In agar disk diffusion test, the zone of inhibition were 25, 9, 10, 10 and 15 mm, respectively. The results showed that the antilisteric activities of essential oils of T. daensis and M. officinalis were superior than that of other essential oils.

Abstract: The inhibitory activity of five bacteriocin (Bac)-producer strains of Staphylococcus aureus was tested against bacteria pathogenic for cattle. Sixty-five epidemiologically unrelated strains of Staph. aureus involved in bovine mastitis were used as indicators in an agar diffusion test. Bacteriocins produced by four strains could inhibit only a limited number of test organisms. However, all 65 indicator strains proved to be susceptible to the combined action of both bacteriocins encoded by pRJ9, a Bac plasmid found in strain A53. Therefore, the bacteriocins produced by this strain may represent new antimicrobial peptides with potential applications in the prevention and treatment of bovine mastitis.

Abstract: The antimicrobial activity of a combination of lactic acid and whey permeate fermented by a nisin-producing Lactococcus lactis strain was tested by the agar diffusion method using bacteria isolated from fish as test organisms. Lactic acid inhibited all bacterial strains studied, but nisin whey permeate inhibited Gram-positive bacteria only. The combination was more effective than lactic acid alone against Pseudomonas fluorescens and Staphylococcus hominis isolated from fish, and Pseudomonas aeruginosa ATCC9721 and Micrococcus luteus ATCC9341.

Abstract: Forty-two isolates of Enterococcus faecalis and 56 isolates of Enterococcus faecium, including 8 vancomycin-resistant strains, were examined for comparative susceptibility to 27 antimicrobial drugs with the agar dilution method, employing Mueller-Hinton (MHA), Iso-Sensitest (ISTA), and Wilkins-Chalgren (WCA) agar. The Bauer-Kirby agar disk diffusion method was used to comparatively test 24 of the agents in parallel. The enterococci yielded better growth on ISTA and WCA. However, WCA completely antagonized co-trimoxazole and, though less, fosfomycin. Importantly, WCA slightly reduced the activities of teicoplanin (minimal inhibitory concentrations, MICs, raised up to twofold) and vancomycin (MICs raised two- to fourfold) against enterococci and staphylococcal quality control strains. Therefore, WCA was judged unsuitable for susceptibility testing of enterococci. For E. faecalis no discrepancies between agar dilution MICs and inhibition zone diameters were encountered with augmentin, ampicillin, ampicillin-sulbactam, chloramphenicol, mupirocin, oxacillin, teicoplanin, and co-trimoxazole. Overall, MHA yielded fewer very major (category I) and major (category II) discrepancies than ISTA. However, numerous minor (category III), slight (category IV), minimal (category V), and/or negligible (category VI) discrepancies were encountered with ciprofloxacin, doxycycline, erythromycin, fosfomycin, fusidic acid, meropenem, ofloxacin and rifampin. With respect to E. faecium, only cefotaxime, mupirocin, oxacillin, and teicoplanin yielded nondiscrepant results. Several very major (I) and major (II) discrepancies were observed with augmentin, ampicillin, ampicillin-sulbactam, doxycycline, fusidic acid, imipenem, and penicillin G. Minor discrepancies (categories III-VI) were particularly numerous with augmentin, chloramphenicol, ciprofloxacin, doxycycline, and piperacillin. The largest numbers of negligible (VI) discrepancies were noted with fosfomycin, fusidic acid, and ofloxacin. It is recommended to test one cephalosporin (cefuroxime or the like) in parallel for educational purposes and to exclude fosfomycin, fusidic acid, and rifampin from test batteries because of the wide scatter of test results. The large number of minimal (V) discrepancies of ciprofloxacin against E. faecalis, the numerous minor (III) and slight (IV) discrepancies of chloramphenicol against E. faecium, and the not insignificant number of very major (I) and minor (III) discrepancies observed with meropenem against isolates of E. faecalis necessitated proposals for new disk intermediate susceptibility criteria.

Abstract: The BIOMIC is a computerized system used to calculate the minimal inhibitory concentration (MIC) of an antimicrobial agent from a zone of inhibition generated by a disk diffusion test. This system was developed using bacterial pathogens of human origin. This study investigated the use of the BIOMIC system for determining MICs for bacterial pathogens from animals. The MICs generated by the BIOMIC system were compared with the MICs generated using a broth microdilution testing method. A total of 663 drug-organism combinations was tested. These combinations included 3 species of gram-positive bacteria, 5 species of gram- negative bacteria, and the antimicrobial agents ampicillin, gentamicin, cephalothin, ciprofloxacin, enrofloxacin, trimethoprim/sulfamethoxazole, tetracycline, and erythromycin. Overall, the MICs generated by the BIOMIC system correlated with the broth microdilution MICs for 72% of the total drug-organism combinations tested. The Pseudomonas aeruginosastrains tested showed the highest agreement between the 2 systems, with 100% for all antibacterial agents tested, whereas Pasteurella haemolytica, Pasteurella multocida,and enterococci showed the least agreement (76%, 57%, and 47%, respectively). Among these organisms, trimethoprim-sulfa showed the least agreement (31%) and ciprofloxacin showed the greatest (91%). These results indicate that the BIOMIC system could be a useful tool in veterinary medicine for producing quantitative antimicrobial suscep- tibility results. However, it is currently unreliable for some drug-bacteria combinations. This discrepancy pos- sibly could be corrected by modification of the software using data points generated by a large-scale study. The importance of antibacterial chemotherapeutic agents in the treatment of infectious disease processes is well established. Historically, a practitioner could select an effective drug based on clinical experience. However, with an increase in bacterial resistance to commonly used antibacterial agents, it has become in- creasingly difficult for clinicians to empirically select an appropriate antibacterial agent. An alternative to empirically selecting an antibacterial agent is in vitro antimicrobial susceptibility testing on bacteria isolated from properly collected samples. There are basically 2 types of results that are provided by in vitro suscep- tibility testing: qualitative results, i.e., susceptible, in- termediate, or resistant, and quantitative results, i.e., the minimum concentration of an antimicrobial agent required to inhibit the growth of the bacteria (MIC). Two common in vitro methods for determining sus- ceptibility profiles of bacterial pathogens are disk dif- fusion testing and dilution testing. Disk diffusion test- ing involves applying a single concentration of an an- timicrobial agent to a seeded agar medium and allow- ing the drug to diffuse into the surrounding medium. The bacteria on the agar medium are thus exposed to a continuous gradient of drug with the concentration diminishing as the distance from the disk increases. A

Abstract: The authors determined the susceptibility of 55 single clinical strains of Helicobacter pylori isolated in the Montreal area to 10 antibiotics by three different methods - an agar dilution technique considered to be the gold standard, a disk diffusion method and the E-test. Testing was performed on Mueller-Hinton agar supplemented with 10% sheep blood; plates were incubated at 37°C for 72 h in a microaerophilic atmosphere. The metronidazole resistance rate is about 11% in the Montreal area. Macrolides are very active against H pylori isolates, with few variations in activity between older and newer molecules. Correlation among different methods was not as good as reported in the literature for metronidazole.

Abstract: A total of 28 traditional drinks derived from 23 plant species in 19 families were tested for their in vitro growth-inhibiting effects against Bifidobacterium adolescents, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus casei, Clostridium perfringens, and Escherichia coil using a paper disc agar diffusion method under anaerobic condition. The responses varied with bacterial strain, plant species and tissue sampled. In a test with C. perfringens at 5 and 10 mg/disc, potent growth inhibition was produced from the extracts of Eucommia ulmoides stems, Pinus densiflora leaves and shoots, Thea sinensis leaves (green and oolong teas) and Zingiber officinale roots. All materials tested did not adversely affect the growth of bifidobacteria, lactobacilli, and E. coli. These results may be an indication of at least one of the pharmacological activities of these plant-derived drinks.

Abstract: OBJECTIVE A comparison of the sensitivity of the agar diffusion method with that of extraction using cell-lines RC-IAL (fibroblastic of rabbit kidney) and HeLa (epithelial carcinoma cells from the cervix uteri of the human uterus), in the in vitro evaluation of materials of medical and hospital. MATERIALS AND METHODS Fifteen samples chosen at random, from among the already known positives and negatives in our stock, were tested and identified as cotton, form, latex, cellulose and acrylic. Besides the samples mentioned, many SDS (GIbco) concentrations were tested experimentally in RC-IAL and HeLa cell cultures. RESULTS Of the 50 samples tested, 44(88%) were positive by both methods. However, when the SDS were compared by using the two methods, positive results were noted in the concentrations of from 0.5 to 0.05 microgram/ml in the agar diffusion ans extraction methods. A cytotoxic effect was only noted in the concentrations of up to 0.25 microgram/ml. CONCLUSION When the SDS was used, differences favorable to the agar diffusion method were observed in the two cell lines, in two concentrations; that is, the sensitivity of this method was quantitatively greater on inspection than that of the extraction method, as well as being the simpler method to use.