Showing papers on "Agar plate published in 2005"

Effect of culture conditions on microorganism identification by matrix-assisted laser desorption ionization mass spectrometry.

Abstract: Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS) has been used to identify bacteria based upon protein signatures. This research shows that while some different proteins are produced by vegetative bacteria when they are cultured in different growth media, positive identification with MALDI-TOF MS is still possible with the protocol established at the Pacific Northwest National Laboratory (K. H. Jarman, S. T. Cebula, A. J. Saenz, C. E. Petersen, N. B. Valentine, M. T. Kingsley, and K. L. Wahl, Anal. Chem. 72:1217-1223, 2000). A core set of small proteins remain constant under at least four different culture media conditions and blood agar plates, including minimal medium M9, rich media, tryptic soy broth (TSB) or Luria-Bertani (LB) broth, and blood agar plates, such that analysis of the intact cells by matrix-assisted laser desorption/ionization mass spectrometry allows for consistent identification.

Isolation and characterization of actinomycetes antagonistic to pathogenic Vibrio spp. from nearshore marine sediments

Abstract: A total of 94 actinomycete strains were isolated from the marine sediments of a shrimp farm, 87.2% belonged to the genus Streptomyces, others were Micromonospora spp. Fifty-one percent of the actinomycete strains showed activity against the pathogenic Vibrio spp. strains. Thirty-eight percent of marine Streptomyces strains produced siderophores on chrome azurol S (CAS) agar plates. Seven strains of Streptomyces were found to produce siderophores and to inhibit the growth of Vibrio spp. in vitro. Two of them belonged to the Cinerogriseus group, the most frequently isolated group of Streptomyces. The results showed that streptomycetes could be a promising source for biocontrol agents in aquaculture.

Characterization of a novel Cr6+ reducing Pseudomonas sp. with plant growth-promoting potential.

Abstract: The isolate RNP4 obtained from a long-term tannery waste contaminated soil was characterized and presumptively identified as Pseudomonas sp. The strain RNP4 tolerated concentrations up to 450 mg Cr6+/L on a Luria-Bartani (LB) agar medium and reduced a substantial amount of Cr6+ to Cr3+ in the LB liquid medium. The ability of performing multifarious activities in tandem suggested the uniqueness of isolate RNP4. The strain produced a substantial amount of indole acetic acid (IAA) in tryptophan-supplemented medium. The strain also exhibited the production of siderophore and solubilization of phosphorus in mineral salt medium and SRS1 medium, respectively. Concurrent production of IAA and siderophore and the solubilization of phosphorus revealed its plant growth promotion potential. Furthermore, the strain was able to promote the growth of black gram, Indian mustard, and pearl millet in the presence of Cr6+. Thus, the innate capability of this novel isolate for parallel bioremediation and plant growth promotion has significance in the management of environmental and agricultural problems.

Use of a selective enrichment broth to recover Clostridium difficile from stool swabs stored under different conditions

Abstract: The recovery of Clostridium difficile from the stools of patients with C. difficile-associated diarrhea was evaluated by use of an enrichment broth (cycloserine-cefoxitin fructose broth supplemented with 0.1% sodium taurocholate [TCCFB]) and was compared to that from selective agar (cycloserine-cefoxitin fructose agar [CCFA]) and alcohol shock followed by inoculation onto blood agar (AS-BA). TCCFB was superior to CCFA and AS-BA, and neither the storage time nor the storage temperature affected the recovery rate.

Production and Validation of the Use of Gamma Phage for Identification of Bacillus anthracis

Abstract: Gamma phage specifically lyses vegetative cells of Bacillus anthracis and serves as part of the basis for identification of isolates from agar cultures. We report our study to standardize gamma phage production and preparation and to validate the assay for routine use. Unstable phage preparations were largely reduced through propagation of phage on blood agar cultures of the avirulent B. anthracis strain CDC684 and were adequately stable for extended storage beyond 1 to 2 years at 4 degrees C, provided that the preparation initially gave rise to clearly discernible plaques (macroplaques, 5 to 10 mm in diameter) on dilution at 1:8 or greater during potency testing with the Sterne strain or its equivalent. The primary intent of the assay was to test nonhemolytic, ground-glass-appearing bacterial B. anthracis-like colonies arising from culture of clinical or nonclinical samples on 5% sheep blood agar. Specifically, the assay was designed to show clear or primarily clear circular zones of lysis on bacterial lawns at the site of gamma phage inoculation after incubation at 35 degrees C +/- 2 degrees C for 20 h. When tested with 51 B. anthracis strains and 49 similar non-B. anthracis Bacillus species, the analytical specificity was >95%, a value that is intentionally low because our study design included two rare nonsusceptible B. anthracis strains as well as a rare susceptible non-B. anthracis strain, B. cereus ATCC 4342. Repeatability, day-to-day precision, and analyst-to-analyst precision were superior. The assay was rugged to variations among phage lots, phage concentration, amounts of bacterial inoculum, and incubation times as short as 6 to 8 h. System suitability evaluation showed improved robustness when bacterial lawns were tested with high- and low-density inoculum using the first and second quadrants of a serial four-quadrant streak on 5% sheep blood agar plates.

Isolation of Candida spp. from mastitic bovine milk in Brazil.

Abstract: The purpose of this study was to isolate yeast (Candida) from the quarter milk of cow udders from 37 dairy farms in Brazil and to identify the different species involved in mastitis. The samples were collected between October 2002 and February 2003. Two-hundred-and-sixty milk samples from cows with clinical and subclinical mastitis were examined. Milk samples were plated onto Blood agar, Mac Conkey agar and Sabouraud dextrose agar. Forty-five (17.3%) samples were positive for the genus Candida. The Candida species isolated were C. krusei (44.5%), C. rugosa (24.5%), C. albicans (8.9%), C. guilliermondii (8.9%), and others (13.2%). We also isolated Escherichia coli (26.5%), coagulase-positive Staphylococcus (25.0%), Streptococcus spp. (8.1%), Enterobacter spp. (8.1%), and other fungi (8.1%), among others.

Comparison of the Quantitative Formalin Ethyl Acetate Concentration Technique and Agar Plate Culture for Diagnosis of Human Strongyloidiasis

Abstract: The quantitative formalin ethyl acetate concentration technique (QFEC) was compared to agar plate culture (APC) for the detection of Strongyloides stercoralis larvae. QFEC could substitute for APC only when the parasite load was higher than 50 larvae per g of stool. This study serves as a good reminder to those conducting stool exams about the sensitivity and specificity limitations of both techniques.

Blood Agar Plates and Hemolysis Protocols

Abstract: In the same discussion, however, they note that Robert Koch preferred plates poured by mixing bacterial inocula with melted gelatin rather than streaking material on the surface. Koch recommended media that were “firm, and where possible, ...transparent...” It appears that pour plates were the standard procedure for many years due largely to problems with surface contamination upon incubation. (It should be noted that, initially, agar “plates” were, indeed, sterilized flat glass plates, not Petrie dishes as we know today.)

Evaluation of Two Chromogenic Agar Media for Recovery and Identification of Staphylococcus aureus Small-Colony Variants

Abstract: To identify the most rapid and reliable technique for recovery and identification of Staphylococcus aureus small-colony variants (SCVs), the colonial appearance of 106 isolates representing SCVs and the normal phenotype were evaluated on two newly described chromogenic agar media. Although almost all of the SCVs grew on the chromogenic agar media, they did not exhibit a change of color. In comparison with conventional media, S. aureus ID agar (SAID; bioMerieux, La Balme Les Grottes, France) showed the most reliable results, with 49 of 53 SCVs tested growing either as an SCV colony or with a normal phenotype after only 24 h of incubation. Growth of SCVs was often not detected before 72 h of incubation on some of the media tested. In conclusion, the most accurate and rapid method to detect both the species S. aureus and the SCV phenotype is to inoculate specimens onto both Columbia blood agar and SAID.

Antibacterial properties of aged dental cements evaluated by direct-contact and agar diffusion tests.

Abstract: Statement of problem Since failure of fixed partial dentures is most frequently caused by caries, it would be advantageous if cements possessed antibacterial properties. Purpose The purpose of this study was to evaluate the antibacterial properties of 3 dental cements using the direct-contact test and agar diffusion test. Material and methods For the direct-contact test, wells (n=4) of microtiter plates were coated with the tested cements (Harvard cement, Duralon, and Ketac-Cem) while Streptococcus mutans suspension was placed directly on the cements. Bacterial growth was evaluated by a temperature-controlled microplate spectrophotometer. Eight wells of bacteria without the tested cements served as the positive control. Six wells of the tested cement without bacteria served as the negative control. For the agar diffusion test, triplicate specimens of freshly mixed cements were poured into uniform wells (5 mm in diameter) punched in the agar plates inoculated with Streptococcus mutans . After incubation at 37°C for 24 hours, the agar plates were examined for bacterial growth and the diameter of the halo formed in the bacterial lawn was measured. In both tests, each cement was mixed in 2 different powder/liquid ratios. For the direct-contact test, data were initially recorded after 1 hour of incubation. Additional experiments were performed on specimens that were aged for 24 hours, 1 week, 1 month, and 3 months before assessment by either direct-contact test or agar diffusion test. The data were subjected to 1-way ANOVA with the Tukey post hoc test (α=.05). Results Compared with the control group, Duralon and Harvard cements demonstrated antibacterial properties even after 3 months with the direct-contact test ( P Conclusions Within the limitations of this study, Duralon and Harvard cements possessed prolonged antibacterial properties, while Ketac-Cem exhibited no antibacterial activity. The direct-contact test may be a more suitable test than the agar diffusion test to evaluate antibacterial properties of definitive cements.

Evaluation of normal ocular bacterial flora with two different culture media.

Abstract: Background: The purpose of this study was to determine if the use of broth culture medium is efficient in investigating bacterial flora of the normal eyelid and conjunctiva. Methods: Samples from the conjunctiva and eyelid of healthy patients of various ages who were undergoing ocular surgeries were obtained and cultured at 3 periods: before topical antibiotic prophylaxis, in the postoperative period during topical antibiotic treatment, and 15 days after discontinuation of antibiotic use. Samples were inoculated into both brain heart infusion broth and blood agar plate, and the growth results of both media were analyzed. Results: Brain heart infusion broth medium showed a significantly higher bacterial growth of gram-positive cocci in most periods. The solid blood agar medium had a higher recovery of gram-positive bacilli before prophylaxis only in the older patients. Interpretation: Our results show that a more complete analysis of eyelid and conjunctival flora can be obtained using both liquid and solid media to increase the chances of isolate recovery. The inclusion of liquid media in this analysis was even more relevant in the period of concomitant use of antibiotic treatment.

Bifidobacterium-selective isolation and enumeration from chicken caeca by a modified oligosaccharide antibiotic-selective agar medium.

Abstract: Aims: To determine the efficacy and selectivity of an acidified, antibiotic-selective, oligosaccharide-containing media for enumerating Bifidobacterium spp. from chicken caeca samples. Methods and Results: Transoligosaccharide propionate agar medium (TOS) modified by addition of mupirocin (50 μg ml−1) and glacial acetic acid (1%, v/v), did not inhibit the growth of bifidobacteria compared with the control media yet inhibited the growth of Lactobacillus acidophilus, Lactobacillus gallinarum, Lactobacillus helveticus and Streptococcus gordonii. Conclusions: Addition of mupirocin (50 μg ml−1) and glacial acetic acid (1%, v/v) to TOS (TOS-AM50), is an effective selective medium for isolation and enumeration of Bifidobacterium spp. from chicken caeca samples. Significance and Impact of the Study: The development of an intestinal bifidobacteria-selective media contributes to the study of probiotics and prebiotics in poultry and potentially other species.

Phenotypic identification of Candida albicans by growth on chocolate agar.

Abstract: In this study, we describe a simple method for the identification of Candida albicans in clinical samples. A total of 383 clinical isolates of Candida species were streaked onto chocolate agar and incubated for 48 h at 37 degrees C in the presence of an atmosphere of 6% CO2. All 208 of the C. albicans isolates tested, developed an easy to identify filamentous colony morphology. Of 175 other Candida species tested, 172 (98.3%) were distinguishable from C. albicans by their smooth colony morphology. Three isolates (1.7%) exhibited weak filamentation after prolonged incubation. Although not a routine medium in medical mycology a significant advantage of using chocolate agar lies in its use in clinical bacteriology laboratories for the isolation of fastidious bacteria. Implementation of the proposed method is applicable across a range of specimen types, thus allowing the direct identification of C. albicans in clinical samples. This simple method may allow a quicker entry into directed treatment.

A Semi-Selective agar medium to detect the presence of Xanthomonas axonopodis pv. malvacearum in naturally infected cotton seed

Abstract: A semi-selective agar medium was developed for detection of Xanthomonas axonopodis pv. malvacearum (Xam) in cotton (Gossypium hirsutum) seed. The basic medium was peptone-sucrose-agar (PSA). Criteria for the semi-selective medium were the typical colony characters of Xam and its pathogenicity on cotton. Several systemic fungicides and antibiotics in different concentrations were tested alone or in combination with others. The final composition of the semi-selective agar medium was established after several attempts in order to inhibit most of the fungal and bacterial saprophytes and favour the development of Xam. It contained PSA + cyclohexamide, cephalexin, pencycuron, triadimenol and tolylfluanid. The bacteria were recovered from naturally infected seeds by the direct plating of 2,000 surface disinfected seeds on the semi-selective medium. The recovery of the pathogen from naturally infected leaf tissues and in dilution plating, on semi-selective medium and on nutrient agar, were comparable. Among the three detection methods tested, the semi-selective medium was found to be the most reliable and quantifiable. Degree of severity of angular leaf spot in the field was not always correlated with the level of infection in the seed. This is the first report of a semi-selective agar medium to detect the presence of Xam in naturally infected cotton seed.

Identification of non-mutans streptococci organisms in dental plaques recovering on mitis-salivarius bacitracin agar medium

Abstract: The objective of this study was to both isolate and identify non-mutans streptococci organisms (non-MSO) from dental plaques recovered on mitis-salivarius sucrose bacitracin agar (MSB) plates. The dental plaque samples, which had been collected from 63 human subjects, were diluted and plated on MSB. The bacteria growing on the MSB plates were then identified with biochemical tests, as well as with 16S rDNA cloning and sequencing techniques. Our data indicated that bacteria from 30 subjects had been recovered on the MSB plates. Among the 21 typical colonies selected from the 30 subjects, 12 colonies, derived from 10 subjects, were identified as non-MSO. These 12 colonies were determined to be Streptococcus anginosus (8 colonies), S. sanguinis (1 colony), and Pantoea agglomerans (3 colonies). These results strongly suggest that a new selective medium will be required for the reliable isolation of mutans streptococci.

The effects of Pseudomonas and Aeromonas strains on Legionella pneumophila growth

Abstract: Manmade water systems such as cooling towers, potable water systems etc. serve as amplifiers for Legionella pneumophila and non-legionella heterotrophic bacteria (NLHB) by providing suitable conditions for growth and multiplication. In this study we investigated the activity of Pseudomonas and Aeromonas strains, which were isolated from water systems of buildings in Istanbul city, on the growth of L. pneumophila serogroup 1 in vitro. It was found that while the cultures of Pseudomonas species (100%) had a stimulating effect on the growth of L. pneumophila, the cultures of Aeromonas species (61.5%) generally inhibited the growth of L. pneumophila on selective agar medium. Also, it was detected that cell-free supernatants (CFSs) of both Aeromonas, except N24 strain, and Pseudomonas species were not capable of inhibiting the growth of L. pneumophila serogroup 1 strains on buffered charcoal yeast extract agar (BCYEA) plates. However, some of these substances were able to stimulate the growth of L. pneumophila on BCYEA without L- Cysteine (UNBCYEA). It can be thought that the growth of L. pneumophila serogroup 1 could be controlled by the NLHB such as Pseudomonas and Aeromonas that live normally in the same water body. The level of the NLHB can change time to time in water systems and this situation can affect outbreaks of Legionnaires disease.

Activity staining method of chitinase on chitin agar plate through polyacrylamide gel electrophoresis

Abstract: A method for detection of chitinase activity on chitin agar plate after polyacrylamide gel electrophoresis is described. Different staining dyes such as calcofluor white M2R, fluorescein isothiocyanate, rhodamine B, ruthenium red and congo red were separately incorporated in chitin agar plates. After running polyacrylamide gel electrophoresis, the gel was transferred onto chitin agar plate containing different dyes for the activity staining. Thin layer of acetate buffer (0.2 M, pH 5) was pored on the gel, which helps faster diffusion of the enzyme from gel onto the plate. After incubation of about 7 h, bands of chitinase were visible by daylight or UV light. The method is very sensitive since it can detect even 0.5 units of chitinase. Thus, this method is sensitive, rapid, user-friendly, reliable and cost effective.

Anti-fungal activity of sulfamethoxazole toward Aspergillus species.

Abstract: Invasive mycosis has significantly increased in frequency among immunocompromised hosts leading to excessive morbidity and mortality. The combination of sulfamethoxazole (SMX) and trimethoprim (TMP) has been used extensively for the treatment and prophylaxis of infections by various microbes. The purpose of this study is to estimate the anti-fungal activity of SMX-TMP and examine the mechanism of activity. To investigate the antimicrobial activity of SMX-TMP in vitro, a mixture of SMX and TMP at 5:1 was serially diluted and added to potato dextrose agar medium or C-limiting agar medium. Aspergillus species were inoculated on the medium plate with SMX-TMP. The growth of A. fumigatus and A. oryzae was inhibited by addition of SMX-TMP. The anti-Aspergillus effect depended on not TMP but SMX and that was inhibited by p-aminobenzoic acid (PABA). A. niger was not sensitive against SMX-TMP in PDA medium, but sensitive in C-limiting medium. Those results showed that the activity depends on culture medium. Furthermore, addition of human serum did not influence the activity of SMX. The finding in this study suggested that SMX might be effective against Aspergillus species in clinical practice and prophylaxis treatment.

A combined agar‐absorption and BIO‐PCR assay for rapid, sensitive detection of Xylella fastidiosa in grape and citrus

Abstract: Application of the polymerase chain reaction (PCR) to disease diagnosis is limited in part by the presence of PCR inhibitors. Inhibition can be overcome and sensitivity increased by culturing bacteria on agar media prior to PCR (termed BIO-PCR). However, Xylella fastidiosa grows slowly, requiring 10–14 days for visible colonies to appear. In this study an agar-absorption BIO-PCR method for detecting X. fastidiosa in grape and citrus plants was developed. Optimum lengths of time for absorption of inhibitors by the agar medium or enrichment of bacteria on the medium were determined for Pierce's disease of grape and citrus variegated chlorosis. When petioles of grape and citrus leaves with symptoms were spotted onto agar media, the spots washed after various time intervals and assayed for X. fastidiosa by real-time PCR, 97% (31 out of 32) and 100% (six out of six) of spots were positive after 2 days and 4 h for grape and citrus, respectively. With direct PCR, only 12·5% (four out of 32) and 33% (two out of six) of spots were positive, respectively, and visible X. fastidiosa colonies were evident after 10 and 14 days, respectively. In a separate experiment with samples from a different vineyard, 46% (13 out of 28) of the grape samples (agar spots) were positive after 1 day and 93% (26 out of 28) after 5 days using agar-absorption PCR. In contrast, all samples were negative by direct PCR. Viable X. fastidiosa were recovered from all samples after 14 days. Further tests with eight randomly selected grape petioles from three Texas vineyards known to have Pierce's disease resulted in 50% being positive by a simple 24 h agar-absorption PCR assay, whereas none was positive by direct PCR. Overall, 10 out of 16 (63%) vines from five vineyards (two in California and three in Texas) were positive after the 24 h agar-absorption PCR assay. In contrast, only one vine was positive by direct PCR. This simple agar absorption-based PCR assay protocol should prove useful for the routine detection of X. fastidiosa and other slow-growing bacteria in the presence of PCR inhibitors.

Phytochemical and antimicrobial properties of extracts of Combretum racemosum

Abstract: Combretum racemosum (P. Beauv.) (Combretaceae), a straggling shrub widespread across Africa is traditionally reputed to be anthelmintic and antimicrobial for genito-urinary and gastrointestinal infections. The methanol and ethyl acetate crude extracts obtained from the whole plant were evaluated invital to determine inhibition of human pathogenic micro organisms made up of five bacteria and three fungi. The extracts inhibited the eight test organisms to different degrees. All the bacteria strains were sensitive to both extracts at concentration ranging from 25 to 125 mg/ml using the agar broth cup diffusion procedure. The sensitivity of Salmonella typhii, Escherica coli and Pseudomonas aeruginosa (gram negative) to both extracts were not concentration dependent, whereas sensitivity of Bacillus subtilis and Staphylococcus aureus (gram positive) were concentration dependent with activity being higher at higher concentrations of ethyl acetate extract. Only the methanol extract exhibited intrinsic antifungal properties on Candida albicans, Asperigillus niger and Dermatophyte sp. with activity comparable to that of the reference drug tioconazole trosyd. Preliminary phytochemical screening of both extracts indicated the presence of alkaloids, steroids, cardiac glycosides, saponins and tannins. INTRODUCTION Combretum racemosum commonly known as Christmas rose, belongs to the family Combretaceae. The plant has been used for several years in African traditional medical practices and as a condiment in soups. It is a shrub indigenous to the tropical and pan tropical regions. In addition to its anthelmintic and antimicrobial properties, the plant is also used for the treatment of haemorrhoids, convulsive coughing, tuberculosis, toothache and male sterility (Burkill, 1985; Oliver-Bever, 1986). Plants belonging to the family Combretaceae are reputed for anthelmintic and antimicrobial activities. Substantial work has been done in this plant family (Adjanohun and Ake Assi, 1972; Bouquet and Debray, 1974; Burkill, 2000; Jossang et al., 1996; Walker, 1953) but there is no report of any antimicrobial study on C. racemosum. In continuation of our studies on biological activities of medicinal plants (Ajaiyeoba et al., 2000; Onocha et al., 2003), we now report on phytochemical and antimicrobial properties of extracts of Combretum racemosum. MATERIALS AND METHODS Collection, Authentication and Extraction of Plant Material The whole plant material of C. racemosum was collected from the University campus and authenticated by Mr. Felix Usang of the Forest Research Institute (FRIN), Ibadan where a voucher specimen was deposited under file number FHI106430. The airdried plant material (whole plant, 946 g) was extracted in hexane, ethylacetate and methanol for 48 hours. On concentration the resultant hexane (18 g), ethylacetate (28 g) and methanol (20 g) extracts were stored in the refrigerator for further use. Proc. WOCMAP III, Vol. 1: Bioprospecting & Ethnopharmacology Eds. J. Bernath, E. Nemeth, L.E. Craker and Z.E.Gardner Acta Hort 675, ISHS 2005 98 Phytochemical Screening Preliminary phytochemical screening for various secondary metabolites such as anthraquinones, tannins, cardiac glycosides, alkaloids, saponin glycosides and the steroidal nucleus were done for the ethyl acetate and methanol plant extracts using the usual procedures (Harborne, 1991). Antimicrobial Assay 1. Microorganisms. Cultures of five human pathogenic bacteria made up of three gram negative and two gram positive bacteria were used for the in vitro antibacterial assay. The species used were Salmonella typhii, Escherica coli, Pseudomonas aeruginosa, Bacillus subtilis and Staphylococcus aureus. For the antifungal assay, three fungi were utilized: Candida albacans, Aspergillus niger, and Dermatophyte sp. All the microorganisms were obtained from the laboratory stock of the department of Microbiology, University of Ibadan, Ibadan. 2. Media. Nutrient broth, nutrient agar, sabourand dextrose agar (SDA), and tryptone soya agar (Oxford Laboratories, UK) were used in the assays. Methanol and ethylacetate (Merck) were also used in solublising the extracts/drugs and as a negative control in the assays. 3. Antimicrobial Agents. Ampicillin, 12.5 μg/ml (Lab Oftalmiso, Spain), and tioconazole cream 1 mg/ml (Pfizer Inc., New York) were included as standard reference drugs in the study. 4. Antimicrobial Activity Determination. The cup agar broth diffusion procedure (Zwadyk, 1972) was used as an overnight broth culture of 1-2 x 10 CFU of each bacterium was used to seed sterile molten agar medium maintained at 45°C. Sterile tryptone soya agar plate was similarly seeded with fungi. When seeded plants had solidified, five wells (10 mm) respectively, were bored in each plate (7 mm, diameter) with an aseptic cork borer when seeded plates had solidified. 200 mg/ml of extract was reconstituted in methanol (or ethyl acetate) and 80 μl dispensed into each of the wells with the aid of a Pastuer pipette. Diameters of zones of inhibition were determined after incubating plates at 37°C for 24 h (bacteria) and at 25°C for 72h (fungi). When seeded with bacteria, each plate had wells filled with methanol (or ethylacetate) as well as ampicillin and for fungi, tioconazole. This method is similar to previous procedures (Kavanagh, 1977). Antimicrobial studies were done in triplicates and diameters of zones of inhibition (mm) are expressed as the mean and standard errors on means. Student’s “T” tests was used to test probability at P< 0.05. RESULTS AND DISCUSSION The results of the phytochemical screening of the hexane, ethylacetate and methanol extracts of the whole plant are presented in Table 1. Preliminary phytochemical screening of all extracts indicated the presence of alkaloids, steroids, cardiac glycosides, saponins and tannins. The antibacterial ties of the ethylacetate and methanol extracts at concentrations ranging from 25 to 125 mg/ml is presented in Table 2. The bacteria used were clinical strains of Salmonella typhii, Escherica coli, Pseudomonas aeruginosa (gram negative), Bacillus subtilis and Staphylococcus aureus (gram positive). Only the ethylacetate and methanol extracts inhibited the 5 test organisms to different degrees. All the bacteria strains were sensitive to both extracts at concentration ranging from 25 to 125 mg/ml using the agar broth cup diffusion procedure. However, the sensitivity of Salmonella typhi, Escherica coli and Pseudomonas aeruginosa (gram negative) to both extracts were not concentration dependent, whereas sensitivity of Bacillus subtilis and Staphylococcus aureus (gram positive) were concentration dependent, activity being higher at higher concentrations of ethyl acetate extract. The result of the antifungal activities of the methanol extract at concentrations ranging from 25 to 125 mg/ml is presented in Table 3. Three clinical strains of human