Ezekiel Green

Bacterial Exopolysaccharides: Functionality and Prospects

Diverse structural, functional and valuable polysaccharides are synthesized by bacteria of all taxa and secreted into the external environment. These polysaccharides are referred to as exopolysaccharides and they may either be homopolymeric or heteropolymeric in composition and of diverse high molecular weights (10 to 1000 kDa). The material properties of exopolysaccharides have revolutionized the industrial and medical sectors due to their retinue of functional applications and prospects. These applications have been extensive in areas such as pharmacological, nutraceutical, functional food, cosmeceutical, herbicides and insecticides among others, while prospects includes uses as anticoagulant, antithrombotic, immunomodulation, anticancer and as bioflocculants. Due to the extensive applications of bacterial exopolysaccharides, this overview provides basic information on their physiologic and morphologic functions as well as their applications and prospects in the medical and industrial sectors.

Inhibitory properties of selected South African medicinal plants against Mycobacterium tuberculosis.

ETHNOPHARMACOLOGICAL RELEVANCE Tuberculosis (TB), caused by Mycobacterium tuberculosis (MTB) is the most commonly notified disease and the fifth largest cause of mortality. One in 10 cases is resistant to treatment in some areas. Several plants are used locally to treat TB-related disease. AIMS OF THE STUDY The aim was to screen selected South African medicinal plants used to treat TB and related symptoms by traditional healers for antimycobacterial activity. MATERIALS AND METHODS Ethnobotanical information on these plants was obtained. Crude acetone, methanol, hexane and ethanol extracts of 21 selected medicinal plants obtained in Venda, South Africa were screened for their ability to inhibit MTB H(37)Ra and a clinical strain resistant to first-line drugs and one second-line drug using tetrazolium microplate assay to determine the minimum inhibitory concentration (MIC). Results were analyzed using Microsoft Excel 2007 and One way ANOVA; p<0.05 was considered for statistical significance. RESULTS Few acetone extracts were active against MTB with MIC under 100 microg/mL. Four plants showed lower MIC values; Berchemia discolor Klotzsch Hemsl 12, 5 microg/mL on H(37)Ra and 10.5 microg/mL on the clinical isolate, Bridelia micrantha Hochst. Baill (25 microg/mL), Warbugia salutaris Bertol. F Chiov (25 microg/mL), and Terminalia sericea Burch ex D. F (25 microg/mL) on both H(37)Ra and clinical isolate. However, the roots of Ximenia caffra Sond. Var. caffra, barks of Sclerocarya birrea (A Rich) Hochst, Asclepias fruticosa L, tubers of Allium sativum L, leaves of Carica papaya L, Solanum panduriforme E. Mey C, and roots of Securidaca longepedunculata Fresen gave MIC greater than 100 microg/mL. CONCLUSION The acetone extracts of Berchemiadiscolor, Bridelia micrantha, Terminalia sericea and Warbugia salutaris could be important sources of mycobactericidal compounds against multidrug-resistant MTB.

Marked susceptibility of South African Helicobacter pylori strains to ciprofloxacin and amoxicillin: Clinical implications

OBJECTIVES Helicobacter pylori-associated infection is common in South Africa, as in other developing countries. Antibiotic resistance is recognised as a major cause of treatment failure. We studied the susceptibility and resistance patterns of H. pylori to guide empiric treatment and prevent the emergence of resistance. METHODS Two hundred H. pylori strains obtained from gastric biopsies of patients presenting with gastric-related morbidities attending Livingstone Hospital, Port Elizabeth, were evaluated for their susceptibility to seven antibiotics - metronidazole, clarithromycin, tetracycline, amoxicillin, gentamicin, ciprofloxacin and erythromycin. H. pylori was isolated following standard microbiology procedures, and susceptibility determined using the Kirby-Bauer disc diffusion and agar dilution methods. Comparisons of antimicrobial resistance rates with sex of the patients were determined using the chi-square test; a p-value of <0.05 was considered significant. RESULTS Marked susceptibility was observed for ciprofloxacin (100%) and amoxicillin (97.5%), and good activity for clarithromycin (80%) and gentamicin (72.5%). However, marked resistance (95.5%) was observed for metronidazole. The minimal inhibitory concentration (MIC) ranged from 0.0625 microg/ml to 8 microg/ml. The lowest MIC, with a range of 0.0625 - 1 microg/ml, was recorded for ciprofloxacin, while the highest (5 - 8 microg/ml) was noted for gentamicin. CONCLUSION Multidrug resistance was commonly encountered - a finding of clinical significance that calls for continuous surveillance of antibiograms to guide empiric treatment. We advocate the inclusion of ciprofloxacin in the treatment regimen of H. pylori infection in our study environment.

Foodborne pathogens recovered from ready-to-eat foods from roadside cafeterias and retail outlets in Alice, Eastern Cape Province, South Africa: public health implications.

This study assessed the microbiological quality of various ready-to-eat foods sold in Alice, South Africa. Microbiological analysis was conducted on 252 samples which included vegetables, potatoes, rice, pies, beef and chicken stew. The isolates were identified using biochemical tests and the API 20E, API 20NE and API Listeria kits; results were analyzed using the one-way-ANOVA test. Bacterial growth was present in all the food types tested; high levels of total aerobic count were observed in vegetables, 6.8 ± 0.07 followed by rice, 6.7 ± 1.7 while pies had the lowest count (2.58 ± 0.24). Organisms isolated included: Listeria spp. (22%), Enterobacter spp. (18%), Aeromonas hydrophila (12%), Klebsiella oxytoca (8%), Proteus mirabilis (6.3%), Staphylococcus aureus (3.2%) and Pseudomonas luteola (2.4%). Interestingly, Salmonella spp. and Escherichia coli were not isolated in any of the samples. There was a statistically significant difference (p < 0.05) in the prevalence of foodborne pathogens from hygienic and unhygienic cafeterias. The results indicated that most of the ready-to-eat food samples examined in this study did not meet bacteriological quality standards, therefore posing potential risks to consumers. This should draw the attention of the relevant authorities to ensure that hygienic standards are improved to curtain foodborne infections.

Selected South African Honeys and Their Extracts Possess In Vitro Anti-Helicobacter pylori Activity

BACKGROUND AND AIMS Eradication of Helicobacter pylori by triple therapy often results in a failure rate of 10-20%; thus, there is a need to seek alternative treatments. The aim of this study was to screen selected South African honeys for their anti-H. pylori activity, to extract the antimicrobial components using organic solvents and to determine the minimum inhibitory concentrations (MICs) of the extracts. METHODS Three locally produced honeys from different regions in South Africa were screened for anti-H. pylori activity at four different concentrations using the agar well diffusion technique. Subsequently, Pure honey was extracted using n-hexane, diethyl ether, chloroform and ethyl acetate; extracts were also examined for anti-H. pylori activity by agar well diffusion method. The MICs of the three most active extracts were determined both by visual inspection and spectrophotometric analysis at 620 nm using the broth microdilution method. The results were analyzed by one-way ANOVA at 95% significance level. RESULTS All honeys demonstrated anti-H. pylori activity and were most active at 75% v/v. The positive control (clarithromycin) recorded a zone diameter of 18.0 ± 7.4 mm not significantly different (p >0.05) from honeys at 75% v/v and solvent extracts. Chloroform extract recorded the lowest MIC(95) values that ranged from 0.156-5% v/v confirming this extract to be the most active. CONCLUSION All honeys demonstrated anti-H. pylori activity at concentrations ≥10%, as did the solvent extracts. Therefore, these honeys and solvent extracts possess potential compounds with therapeutic activity that could be further exploited as lead molecules in the treatment of H. pylori infections.

Bioflocculant production by a consortium of Streptomyces and Cellulomonas species and media optimization via surface response model

Species of actinobacteria previously isolated from Tyume River in the Eastern Cape Province of South Africa and identified by 16S rDNA sequence as Cellulomonas and Streptomyces species were evaluated as a consortium for the production of bioflocculant. Sucrose, peptone and magnesium chloride were the nutritional sources which supported optimal production of bioflocculant resulting in flocculation activities of 91%, 82% and 78% respectively. Response surface design revealed sucrose, peptone and magnesium chloride as critical media components following Plackett-Burman design, while the central composite design showed optimum concentration of the critical nutritional source as 16.0 g/L (sucrose), 1.5 g/L (peptone) and 1.6g/L (magnesium chloride) yielding optimal flocculation activity of 98.9% and bioflocculant yield of 4.45 g/L. FTIR spectrometry of the bioflocculant indicated the presence of carboxyl, hydroxyl and amino groups, typical for heteropolysaccharide, while SEM imaging revealed an interwoven clump-like structure. The molecular weight distribution of the constituents of the bioflocculants ranged 494.81-18,300.26 Da thus, an indication of heterogeneity in composition. Additionally, the chemical analyses of the purified bioflocculant revealed the presence of polysaccharides and proteins with neutral sugar, amino sugar and uronic acids in the following concentration: 5.7 mg, 9.3mg and 17.8 mg per 100mg. The high flocculation activity of the bioflocculant suggests commercial potential.

A Freshwater Streptomyces, Isolated from Tyume River, Produces a Predominantly Extracellular Glycoprotein Bioflocculant

We evaluated bioflocculant production by a freshwater actinobacteria whose 16S rDNA nucleotide sequence was deposited in GenBank as Streptomyces sp. Gansen (accession number HQ537129). Optimum culture conditions for bioflocculant production were an initial medium pH of 6.8, incubation temperature of 30 °C, agitation speed of 160 rpm and an inoculum size of 2% (v/v) of cell density 1.5 × 108 cfu/mL. The carbon, nitrogen and cation sources for optimum bioflocculant production were glucose (89% flocculating activity), ammonium sulfate (76% flocculating activity) and MgCl2. Bioflocculant pyrolysis showed three step decomposition indicative of three components while chemical analyses showed 78% carbohydrate and 22% protein (wt/wt). The mass ratio of neutral sugar, amino sugar and uronic acids was 4.6:2.4:3. FTIR spectrometry indicated the presence of carboxyl, hydroxyl and amino groups, typical for heteropolysaccharide. The bioflocculant showed a lattice structure as seen by SEM imaging. Its high flocculation activity suggests its suitability for industrial applicability.

An African perspective on Helicobacter pylori: prevalence of human infection, drug resistance, and alternative approaches to treatment

Abstract Helicobacter pylori is a Gram-negative, micro-aerophilic, motile, curved rod that inhabits the gastric mucosa of the human stomach. It chronically infects thousands of millions of people world-wide, and is one of the most genetically diverse of bacterial species. Infection with the bacterium leads to chronic gastritis, peptic ulceration, gastric cancers and gastric mucosa-associated lymphoid-tissue (MALT) lymphoma. The prevalence of infection appears to be partly determined by geographical and socio–demographic factors, being higher in Africa than elsewhere. Current treatment, based on potent combinations that each consist of a proton-pump inhibitor and two antibiotics, is successful in 80%–90% of patients. Some undesirable side-effects, poor patient compliance and drug resistance are, however, associated with significant levels of treatment failure and with contra-indications for some patients. Antibiotic resistance in H. pylori is a growing global concern that merits the urgent attention of public-health authorities. Numerous pieces of clinical evidence have revealed that eradication of the organism from a patient results in improvement of gastritis and drastically decreases the frequency of relapse of gastric and duodenal ulcers. Natural products, including medicinal plants and honey, may offer useful alternatives in the treatment of H. pylori-related infections.

Volatile compounds in the stem bark of Sclerocarya birrea (Anacardiaceae) possess antimicrobial activity against drug-resistant strains of Helicobacter pylori

The aim of this study was to isolate and identify phytochemicals with anti-Helicobacter pylori activity from the stem bark of Sclerocarya birrea. The plant crude extract was fractionated by silica gel column and thin layer chromatography techniques, initially with ethyl acetate (EA) and subsequently with a combination of ethyl acetate/methanol/water (EMW). Further fractionation and identification of the phytoconstituents was achieved by gas chromatography and mass spectrometry (GC/MS) analysis. The antimicrobial activity of the fractions and compounds was evaluated against five metronidazole- and clarithromycin-resistant strains of H. pylori as well as a reference strain ATCC 43526 using the microbroth dilution technique. Amoxicillin was included in the experiments as a positive control antibiotic. Of the 18 fractions collected, 16 demonstrated anti-H. pylori activity with 50% minimum inhibitory concentration (MIC(50)) values ranging from 310 μg/mL to 2500 μg/mL. Two of the fractions (EMW fraction 6 and EA fraction 1) revealed the presence of 5 and 24 compounds, respectively, representing 40.5% and 86.57% of the total composition. Most of the compounds were essential oils, with terpinen-4-ol being the most abundant agent (35.83%), followed by pyrrolidine (32.15%), aromadendrene (13.63%) and α-gurjunene (8.77%). MIC(50) ranges for amoxicillin, terpinen-4-ol and pyrrolidine were 0.0003-0.06 μg/mL, 0.004-0.06 μg/mL and 0.005-6.3 μg/mL, respectively. The inhibitory activities of terpinen-4-ol and pyrrolidine were similar to amoxicillin (P>0.05). Most of these compounds are being reported in this plant for the first time and may represent new sources of therapeutically useful compounds against H. pylori.

Helicobacter pylori Antigenemia in an Asymptomatic Population of Eastern Cape Province, South Africa: Public Health Implications

Helicobacter pylori is a major cause of such upper gastrointestinal diseases as gastritis, peptic ulcer, and gastric cancer. The risk of infection is increased in those living in the developing world, which has been ascribed to precarious hygiene standards, crowded households, and deficient sanitation common in this part of the world. Fecal samples were collected from 356 apparently healthy subjects, consisting of 168 males and 188 females aged from 3 months to > or = 60 years (Mean = 31 years). A standardized questionnaire describing demographic characteristic including age, sex, household hygiene, socioeconomic status, and so on was applied. A sandwich-type enzyme immunoassay amplification technology (Amplified IDEIA Hp StAR, Oxoid, UK) was used to analyze the fecal samples for the detection of H. pylori antigens using monoclonal antibodies specific for H. pylori antigens. Fishers exact test was used to assess the univariate association between H. pylori infection and the possible risk factors. Odds ratio (OR) and the corresponding 95% confidence interval (CI) were calculated to measure the strength of association using EPI INFO 3.41 package. P values of < .05 were required for significance. H. pylori antigen was detected in 309 of the 356 subjects giving an overall prevalence of 86.8%. Prevalence increased with age from 75.9% in children < 12 years age to 100% in young adults aged 25-47 years and subjects aged > or = 60 years (P < .05). H. pylori prevalence was higher in females than in males. Of 188 females who participated in the study, H. pylori antigen was detected in 172 (91.5%) versus 144 (85.7%) 168 males (P > .05). Interestingly, H. pylori antigen was detected more often (100%) in the high socioeconomic group than in those of low socioeconomic group (85.9%) (P > .05). The results of this study have revealed a high prevalence of H. pylori antigens in fecal samples of asymptomatic individuals in the Nkonkobe municipality, an indication of active infection. Socioeconomic status, contaminated water, and poor sanitation may play a role in H. pylori transmission in this population. This finding is of public health and epidemiologic significance.