Nicoline F. Tanih

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.

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.

Molecular Detection of Antibiotic Resistance in South African Isolates of Helicobacter pylori

Rapid diagnosis and treatment of Helicobacter pylori (H. pylori) presents a challenge. We aimed at investigating the presence of H. pylori, susceptibility profile, and associated mutations in an effort to validate the effectiveness of GenoType HelicoDR assay in H. pylori typing in our environment. Two hundred and fifty-four biopsy specimens were cultured and DNA extracted from seventy-eight positive cultures using the Qiagen DNA extraction kit. The GenoType Helico DR which employs reverse hybridisation was used to confirm the presence of H. pylori, determination of its susceptibility to antimicrobials, and detection of mutations conferring resistance to clarithromycin and fluoroquinolones. The organism was isolated from 168/254 (66.1 %) of the specimens by culture. Of the 78 strains used for further investigation, 12/78 (15.38%) were resistant to clarithromycin while 66/78 (84.61%) were susceptible. For fluoroquinolone, 70/78 (89.74%) strains were susceptible while 8 (10.26%) were resistant. Mutations were observed in 17 strains with A2147G being the most prevalent; A2146C and D91N were the least. The reverse hybridisation assay is an easy and fast technique in confirming the presence of H. pylori, its antimicrobial profile, and associated mutations. Analysis regarding the suitability of this assay for H. pylori typing is warranted in other regions.

Helicobacter pylori prevalence in dyspeptic patients in the Eastern Cape province - race and disease status

Objectives. We examined Helicobacter pylori infection in patients with gastric-related morbidities at Livingstone Hospital, Port Elizabeth, to determine the prevalence and risk factors for infection according to race, endoscopic diagnosis, age and sex. Methods. Gastric biopsies were collected from 254 consecutive patients and H. pylori isolated on Columbia agar base supplemented with 7% sheeps blood and Skirrows supplement containing trimethoprim (2.5 mg), vancomycin (5 mg) and cefsulodin (2.5 mg). Amphotericin (2.5 mg) was added to the medium. Recovered isolates were identified following standard microbiology and biochemical techniques. Presumptive isolates were further confirmed by polymerase chain reaction (PCR) targeting the glmM gene. Fishers exact test was used to assess the univariate association between H. pylori infection and the possible risk factors. Odds ratios (ORs) and corresponding 95% confidence intervals (CIs) were calculated to measure the strength of association, using EPI INFO 3.41 software. P-values <0.05 were required for significance. Results. The overall prevalence of H. pylori was 66.1% (168/254). Of the 168 positive subjects, H. pylori prevalence was highest in patients with non-ulcer dyspepsia (NUD) (32.7%; 55/168), and lowest (0%; 0/168) in those with atypical oesophageal reflux disease and gastroduodenitis, respectively. The prevalence of infection was highest among coloureds (68.4%; 89/130) and lowest in whites (59.5%; 25/ 42). Prevalence increased with age. Conclusion. The prevalence of H. pylori is high in dyspeptic patients in Eastern Cape Province. Gender, antibiotic treatment and alcohol consumption may be risk factors for infection. These findings are of clinical and epidemiological significance.

Prevalence of Helicobacter pylori vacA, cagA and iceA genotypes in South African patients with upper gastrointestinal diseases.

Clinical response to Helicobacter pylori infection may be determined by specific virulence-associated genotypes which varies geographically. The aim of this study was to investigate the diversity of putative virulence markers of H. pylori; cagA, vacA and iceA in the Eastern Cape Province of South Africa. One hundred H. pylori strains obtained from dyspeptic patients were used. Gastric biopsies were obtained from 254 dyspeptic patients. H. pylori was cultured and strains were studied. Bacterial genotypes cagA, vacA (s and m subtypes) and iceA were analysed by PCR using specific primers. CagA was identified in 90% of the strains investigated. Fifty-eight of the 100 strains had the vacA signal sequence genotype s1 and 26 had subtype s2. Combined vacA s1/s2 was detected in 16 of the strains. VacA middle region analysis showed that 8 (8%) strains were m1 while 50 were m2. Combined vacA m1/m2 was detected in 36 of the strains. s1m2 (20%) and s2m2 (20%) genotypes were the most common allelic combinations of the vacA gene among the strains. Multiple vacA genotypes were detected in this study. Twenty-six percent of the strains identified had both iceA1 and iceA2. All our strains tested positive for the ureC (glmM) gene. This study reveals a high prevalence of vacA, cagA and iceA2.

The Acetone Extract of Sclerocarya birrea (Anacardiaceae) Possesses Antiproliferative and Apoptotic Potential against Human Breast Cancer Cell Lines (MCF-7)

Interesting antimicrobial data from the stem bark of Sclerocarya birrea, which support its use in traditional medicine for the treatment of many diseases, have been delineated. The current study was aimed to further study some pharmacological and toxicological properties of the plant to scientifically justify its use. Anticancer activity of water and acetone extracts of S. birrea was evaluated on three different cell lines, HT-29, HeLa, and MCF-7 using the cell titre blue viability assay in 96-well plates. Apoptosis was evaluated using the acridine orange and propidium iodide staining method, while morphological structure of treated cells was examined using SEM. The acetone extract exhibited remarkable antiproliferative activities on MCF-7 cell lines at dose- and time-dependent manners (24 h and 48 h of incubation). The extract also exerted apoptotic programmed cell death in MCF-7 cells with significant effect on the DNA. Morphological examination also displayed apoptotic characteristics in the treated cells, including clumping, condensation, and culminating to budding of the cells to produce membrane-bound fragmentation, as well as formation of apoptotic bodies. The acetone extract of S. birrea possesses antiproliferative and apoptotic potential against MCF-7-treated cells and could be further exploited as a potential lead in anticancer therapy.

Increasing trend of metronidazole resistance in the treatment of Helicobacter pylori infection: A global challenge

Helicobacter pylori are gram negative spiral bacteria that colonize the human stomach. Infection with H. pylori is associated with chronic gastritis, peptic ulcer, gastric adenocarcinoma and gastric mucosaassociated lymphoid tissue (MALT) lymphoma. Antibiotic resistance is an ever increasing problem with the treatment of most microbial infections including H. pylori; and has become a growing problem worldwide with the eradication of this organism. In recent years, several treatment regimens have been proposed for H. pylori eradication. However, the only conditions for which such treatment is strongly recommended on the basis of unequivocal supporting evidence are peptic ulcer disease and low grade gastric MALT lymphoma. Success of antimicrobial regimens for H. pylori eradication depends on patient compliance and lack of antimicrobial resistance. Metronidazole (Mtz) containing regimens have been shown to limit effectiveness because of increasing prevalence of resistance to this drug. A high prevalence (> 90%) of Mtz resistance in H. pylori has been reported especially in developing countries. Mtz resistance may be mediated through an inability of Mtz-resistant strains to remove oxygen from the site of Mtz reduction, thereby preventing Mtz activation. This has been attributed to a mutation on the frxA and/or rdxA genes resulting in strains of the organism with defective nitro-reductases coded by these genes. Infection by Mtz or amoxicillin resistant strains is an important factor leading to treatment failure; subjecting all H. pylori clinical isolates to susceptibility testing most especially to Mtz is recommended. If not possible, a program to survey the prevalence of resistance should be implemented in a given area or population. This increasing emergence of antimicrobial resistance in H.pylori treatment posses serious public health problems and is therefore necessary that new drug regimens be examined.

Helicobacter pylori infection and transmission in Africa: Household hygiene and water sources are plausible factors exacerbating spread

Helicobacter pylori (H. pylori) is a microaerophilic motile curve rod that inhabits the gastric mucosa of the human stomach. The organism chronically infects billions of people worldwide and is one of the most genetically diverse of bacterial species. Infection with the bacterium which leads to chronic gastritis, peptic ulceration, gastric cancers and gastric malt lymphoma has been reported to follow a pattern linked to geographic and socio-demographic factors. Studies have documented a higher prevalence in Africa than elsewhere although the pathological outcomes do not correlate with infection. H. pylori transmission pathways are still vague, but the risks of transmission include precarious hygiene standards, over-crowding and contaminated environment and water sources amongst others. The possible routes of transmission include oral-oral, faecal-oral and person- to -person, either with or without transitional transmission steps during episodes of diarrhoea or gastro-oral contact in the event of vomiting. Use of contaminated water including municipal tap water has also been suspected to have a high impact in the transmission of the organism. To generate the data presented in this paper, we conducted an internet based search on relevant literature pertaining to H. pylori epidemiology in general and Africa in particular. Sites such as Pubmed, AJOL, Scopus and Goggle scholar were mainly used. This paper therefore attempts to appraise the role of household hygiene and water sources in the transmission of this organism in the developing world context.

Characterisation of the genes encoding resistance to metronidazole (rdxA and frxA) and clarithromycin (the 23S-rRNA genes) in South African isolates of Helicobacter pylori

Abstract Helicobacter pylori has been incriminated in human diseases, such as peptic ulcer, gastritis and gastric malignancy. Although modern triple-drug regimens are usually highly effective in the treatment of H. pylori infection, the emergence of resistance to two of the most used antibiotics, metronidazole (Mtz) and clarithromycin (Cla), is a serious and increasing problem. Truncations in the rdxA and frxA genes of H. pylori are thought to be associated with Mtz resistance whereas mutations in the pathogen’s 23S-ribosomal-RNA (23S-rRNA) genes are associated with Cla resistance. In a recent study, PCR and sequence analysis of the rdxA, frxA and 23S-rRNA genes were used to explore the genetic basis of resistance to Mtz and Cla in H. pylori. When 200 isolates of H. pylori from the Eastern Cape province of South Africa were tested for antibiotic susceptibility, almost all (95·5%) were found resistant to Mtz and 20·0% were found resistant to Cla. Only the Mtz-resistant isolates showed rdxA and frxA truncation. Two point mutations were detected in the 23S-rRNA genes of the Cla-resistant isolates. Many significant changes (resulting in 13 amino-acid substitutions in nine loci and truncated proteins in 14 loci) were observed in the rdxA genes of the Mtz-resistant isolates, and it appears that, compared with the rarer changes detected in frxA, such mutations may contribute more significantly to the high prevalence of Mtz resistance. To guide empiric treatment, the genotypes and antibiotic susceptibility of H. pylori in the Eastern Cape province of South Africa need to be monitored regularly.