David A. Akinpelu

Interactions of Antibiotics and Methanolic Crude Extracts of Afzelia Africana (Smith.) Against Drug Resistance Bacterial Isolates

Infection due to multidrug resistance pathogens is difficult to manage due to bacterial virulence factors and because of a relatively limited choice of antimicrobial agents. Thus, it is imperative to discover fresh antimicrobials or new practices that are effective for the treatment of infectious diseases caused by drug-resistant microorganisms. The objective of this experiment is to investigate for synergistic outcomes when crude methanolic extract of the stem bark of Afzelia africana and antibiotics were combined against a panel of antibiotic resistant bacterial strains that have been implicated in infections. Standard microbiological protocols were used to determine the minimum inhibitory concentrations (MICs) of the extract and antibiotics, as well as to investigate the effect of combinations of the methanolic extract of A. africana stem bark and selected antibiotics using the time-kill assay method. The extract of Afzelia africana exhibited antibacterial activities against both Gram-negative and Gram-positive bacteria made up of environmental and standard strains at a screening concentration of 5 mg/mL. The MICs of the crude extracts and the antibiotics varied between 1 μg/mL and 5.0 mg/mL. Overall, synergistic response constituted about 63.79% of all manner of combinations of extract and antibiotics against all test organisms; antagonism was not detected among the 176 tests carried out. The extract from A. africana stem bark showed potentials of synergy in combination with antibiotics against strains of pathogenic bacteria. The detection of synergy between the extract and antibiotics demonstrates the potential of this plant as a source of antibiotic resistance modulating compounds.

Biocidal activity of partially purified fractions from methanolic extract of Garcinia kola (Heckel) seeds on bacterial isolates

The in vitro antibacterial activity of crude methanolic extract of the seeds of Garcinia kola was investigated. The extracts exhibited antibacterial activities with zones of inhibition ranging from 10 mm to 25 mm. The minimum inhibitory concentration of the diethyl ether fraction was between 0.313 and 5.0 mg/ml, while that of butanol fraction varied from 0.157 to 5.0 mg/ml. The butanol fraction killed about 77% of Bacillus anthracis and 79% of Escherichia coli cells within 120 min at a concentration of 5.0 mg/ml. Protein leakage from the B. anthracis and E. coli cells when exposed to the butanol and diethyl ether fractions was observed. We conclude that Garcinia kola seed extract has a broad spectrum antibacterial activity, with the butanol and diethyl ether fractions being bactericidal as exemplified by the killing rate and protein leakage regimes, which suggest cell membrane disruption as a mechanism of action of the extract.

Antibiotic Producing Potentials of Three Freshwater Actinomycetes Isolated from the Eastern Cape Province of South Africa

Crude extracts of three actinomycetes species belonging to Saccharopolyspora (TR 046 and TR 039) and Actinosynnema (TR 024) genera were screened for antibacterial activities against a panel of several bacterial strains. The extracts showed antibacterial activities against both gram-negative and gram-positive test bacteria with inhibition zones ranging from 8 to 28 mm (TR 046); 8 to15 mm (TR 039); and 10 to 13 mm (TR 024). The minimum inhibitory concentrations ranged from 0.078 to 10 mg/mL (TR 046); 5 to >10 mg/mL (TR 039); and 1.25 to 5 mg/mL (TR 024). Time-kill studies revealed that crude extract of TR 046 showed strong bactericidal activity against Bacillus pumilus (ATCC14884), reducing the bacterial load by 104 cfu/mL and 102 cfu/mL at 4× MIC and 2× MIC, respectively, after 6 h of exposure. Similarly, against Proteus vulgaris (CSIR 0030), crude extract of TR 046 achieved a 0.9log10 and 0.13log10 cfu/mL reduction at 5 mg/mL (4× MIC) and 1.25 mg/mL (2× MIC) after 12 h of exposure. The extract was however weakly bactericidal against two environmental bacterial strains (Klebsiella pneumoniae and Staphylococcus epidermidis); and against Pseudomonas aeruginosa (ATCC 19582): the extract showed bacteriostatic activities at all concentrations tested. These freshwater actinomycetes appear to have immense potential as a source of new antibacterial compound(s).

Preliminary Phytochemical Screening and Antibacterial Properties of Crude Stem Bark Extracts and Fractions of Parkia biglobosa (Jacq.)

A methanolic crude extract of Parkia biglobosa was prepared and later partitioned in succession with different solvents of increasing polarity ranging from n-hexane, chloroform and ethyl acetate to butanol. Phytochemical screening of the extract revealed the presence of alkaloids, tannins, saponins, flavonoids, steroids, glycoside and sugars. The inhibition zones exhibited by the extract against the tested bacteria ranged between 14 ± 0.00 mm (against Escherichia coli) and 28 ± 0.71 mm (against Pseudomonas aeruginosa). The MIC of the methanolic extract of P. biglobosa against isolates ranged between 0.63 mg/mL and 5 mg/mL, while the MIC values exhibited by the n-hexane and aqueous fractions ranged between 0.63 mg/mL and 10 mg/mL. Overall the extract and fractions of P. biglobosa used in this work were found to possess antimicrobial properties which compared favourably with those of streptomycin. These observations make this plant a potential source of bioactive compounds that can be used in management of bacterial infections. The use of this plant as herbal medicaments in African countries and the reports on the toxicity of the plant further show that the plant is non-toxic to humans.

Studies on the biocidal and cell membrane disruption potentials of stem bark extracts of Afzelia africana (Smith)

We had recently reported antibacterial activity in the crude extract of the stem bark of Afzelia africana (Akinpelu et al., 2008). In this study, we assessed the biocidal and cell membrane disruption potentials of fractions obtained from the crude extract of the plant. The aqueous (AQ) and butanol (BL) fractions exhibited appreciable antibacterial activities against the test bacteria. The minimum inhibitory concentrations of the AQ and BL fractions ranged between 0.313 and 2.5 mg/ml, while their minimum bactericidal concentrations varied between 0.625 and 5.0 mg/ml. Also, the AQ fraction killed about 95.8% of E. coli cells within 105 min at a concentration of 5 mg/ml, while about 99.1% of Bacillus pumilus cells were killed by this fraction at the same concentration and exposure time. A similar trend was observed for the BL fraction. At a concentration of 5 mg/ml, the butanol fraction leaked 9.8 microg/ml of proteins from E. coli cells within 3 h, while the aqueous fraction leaked 6.5 microg/ml of proteins from the same organisms at the same concentration and exposure time. We propose that the stem bark of Afzelia africana is a potential source of bioactive compounds of importance to the pharmaceutical industry.

In-Vitro Antibacterial Properties of Crude Aqueous and n-Hexane Extracts of the Husk of Cocos nucifera

The increasing numbers of cases of antibiotic resistance among pathogenic bacteria such as Vibrio species poses a major problem to the food and aquaculture industries, as most antibiotics are no longer effective in controlling pathogenic bacteria affecting these industries. Therefore, this study was carried out to assess the antibacterial potentials of crude aqueous and n-hexane extracts of the husk of Cocos nucifera against some selected Vibrio species and other bacterial pathogens including those normally implicated in food and wound infections. The crude extracts were screened against forty-five strains of Vibrio pathogens and twenty-five other bacteria isolates made up of ten Gram positive and fifteen Gram negative bacteria. The aqueous extract was active against 17 of the tested bacterial and 37 of the Vibrio isolates; while the n-hexane extract showed antimicrobial activity against 21 of the test bacteria and 38 of the test Vibrio species. The minimum inhibitory concentrations (MICs) of the aqueous and n-hexane extracts against the susceptible bacteria ranged between 0.6–5.0 mg/mL and 0.3–5.0 mg/mL respectively, while the time kill study result for the aqueous extract ranged between 0.12 Log10 and 4.2 Log10 cfu/mL after 8 hours interaction in 1 × MIC and 2 × MIC. For the n-hexane extract, the log reduction ranged between 0.56 Log10 and 6.4 Log10 cfu/mL after 8 hours interaction in 1 × MIC and 2 × MIC. This study revealed the huge potential of C. nucifera extracts as alternative therapies against microbial infections.

Evaluation of Antibacterial and Antifungal Properties of Alchornea laxiflora (Benth.) Pax. & Hoffman

Alchornea laxiflora leaf extract was tested against a range of microorganisms using standard microbiological methods for antimicrobial activities. The extract inhibited the growth of all the bacterial and 15 fungal isolates tested. The zones of inhibition exhibited against the test bacteria ranged between 12 mm and 24 mm and between 11 mm and 24 mm for the extract and the antibiotic streptomycin, respectively. The zones of inhibition observed against the fungal isolates by the extract ranged between 12 mm and 23 mm. The minimum inhibitory concentrations (MICs) and the minimum bactericidal concentrations (MBCs) exhibited by the extract against test bacteria ranged between 0.78 mg/mL–25 mg/mL and 1.56 mg/mL–25 mg/mL, respectively, while the MICs and minimum fungicidal concentrations (MFCs) values for the test fungi ranged between 8.75 mg/mL–35.00 mg/mL and 8.75 mg/mL–35.00 mg/L, respectively. The preliminary phytochemical screening of the extract revealed the presence of alkaloids, tannins, flavonoids, saponins, and reducing sugars as major phytoconstituents in the extract. A. laxiflora leaf extract is a potent source of antibacterial and antifungal compounds; further studies on the extract are ongoing in our laboratories to elucidate the probable mechanism(s) of action on bacteria and fungi found to be susceptible to the extract.

Stem bark extract and fraction of Persea americana (Mill.) exhibits bactericidal activities against strains of bacillus cereus associated with food poisoning.

The study investigates the in vitro antibacterial potentials of stem bark extracts of Persea americana on strains of Bacillus cereus implicated in food poisoning. The crude stem bark extracts and butanolic fraction at a concentration of 25 mg/mL and 10 mg/mL, respectively, exhibited antibacterial activities against test isolates. The zones of inhibition exhibited by the crude extract and the fraction ranged between 10 mm and 26 mm, while the minimum inhibitory concentration values ranged between 0.78 and 5.00 mg/mL. The minimum bactericidal concentrations ranged between 3.12 mg/mL–12.5 mg/mL and 1.25–10 mg/mL for the extract and the fraction, respectively. The butanolic fraction killed 91.49% of the test isolates at a concentration of 2× MIC after 60 min of contact time, while a 100% killing was achieved after the test bacterial cells were exposed to the butanolic fraction at a concentration of 3× MIC after 90 min contact time. Intracellular protein and potassium ion leaked out of the test bacterial cells when exposed to certain concentrations of the fraction; this is an indication of bacterial cell wall disruptions by the extract’s butanolic fraction and, thus, caused a biocidal effect on the cells, as evident in the killing rate test results.

In vitro antilisterial properties of crude aqueous and nhexane extracts of the husk of Cocos nucifera

The in vitro antilisterial activities and time kill regimes of crude aqueous and n-hexane extracts of the husk fiber of Cocos nucifera were assessed. The aqueous extracts were active against 29 of the 37 test Listeria isolates while the n-hexane extracts were active against 30. The minimum inhibitory concentrations (MICs) of all the susceptible bacteria ranged between 0.6 and 2.5 mg/ml for the aqueous fraction and between 0.6 and 5.0 mg/ml for the n-hexane extract. The average log reduction in viable cell count in the time kill assay ranged between 0.32 Log 10 and 3.2 Log 10 cfu/ml after 4 h of interaction, and between 2.6 Log 10 and 4.8 Log 10 cfu/ml after 8 h interaction in 1 × MIC and 2 × MIC (aqueous extract); and between 2.8 Log 10 and 4.8 Log 10 cfu/ml after 4 h of interaction, and 3.5 Log 10 to 6.2 Log 10 cfu/ml after 8 h interaction in 1 × MIC and 2 × MIC for the n-hexane extract. The extract was bactericidal against one of the test bacteria at 1 × MIC and against three of the test bacteria at 2 × MIC for the 8 h interaction period for the aqueous extract, while for the n-Hexane fraction; the extract was bactericidal against all the five test bacteria at both MICs after the 8 h interaction period. We suggested that the crude aqueous and n-hexane extracts of the husk of C. nucifera could be bacteriostatic or bactericidal depending on the time of exposure and concentration. Key words: Cocos nucifera, n-hexane extract, aqueous extract, minimum inhibitory concentration, time-kill.