Ethno-medicinal documentation of polyherbal medicines used for the treatment of tuberculosis in Amathole District Municipality of the Eastern Cape Province, South Africa.

CONTEXT: Tuberculosis (TB) has remained a devastating global public health problem. In the continuing search for effective treatment, polyherbal remedies used as alternative medicines in the Eastern Cape Province of South Africa were surveyed. OBJECTIVE: The survey collected information and documents the list of ingredients such as the name of the plants used including the non-herbal inclusions, type and dosage of polyherbal formulations used for the treatment of TB. MATERIALS AND METHODS: The survey was conducted over a period of 6 months using semi-structured questionnaires amidst informal conversations with the traditional healers in five communities in the study area. The chosen study area is the third infected Province with TB in South Africa. RESULTS: A total of nine polyherbal preparations were collected. Information on the parts of the plant used, mode of preparation and the dosage used were documented. In total, the herbs belong to 20 families of which Apiaceae, Liliaceae, Strychnaceae, Rutaceae and Hypoxidaceae are the most prominent. However, members of Apiaceae were commonly mentioned for the preparation of the remedies. The two most frequently used plants were Allium sativum L. (Liliaceae) and Strychnos decussata (Pappe) Gilg. (Strychnaceae). Rhizome was the commonest parts used, followed by the roots and barks. CONCLUSIONS: This paper provides significant ethno-medicinal information on polyherbal medicines used for the treatment of TB in the study area. The therapeutic claims made on medicinal plants used for the preparations are well supported by the literature, with many of the species having antimicrobial properties.

Inhibitory and bactericidal activity of selected South African honeys and their solvent extracts against clinical isolates of Helicobacter pylori.

The growing problem of antibiotic resistance by Helicobacter pylori demands the search for novel compounds, especially from natural sources. We evaluated the anti-H. pylori activity of six local honeys at different concentrations as well as their solvent extracts by the Hole Plate diffusion method. The minimum inhibitory concentration (MIC50) of the two most active extracts of each honey was determined by the broth microdilution method; and the time kill assay of the most active extract of each honey determined by viability studies. Data were analyzed by one-way ANOVA test at 95% significance level. All the honey varieties as well as their solvent extracts demonstrated varying levels of antibacterial activity based on different mean zone diameters [16.0mm (crude) to 22.2mm (extract)] and percentage susceptibilities [73.3% (crude) to 93.3% (extract)] of the test isolates. The chloroform extracts of Pure Honey (PH) and Champagne Royal Train (CRT) recorded MIC50 ranges of 0.01-10% and 0.625-10 % (v/v) respectively; that were not significantly different (P > 0.05) from amoxicillin (0.001-1.25mg/mL), the positive control. The most potent bactericidal effect against the test isolates was obtained with 5% v/v (1/2 MIC) concentration of chloroform extract of PH from 42-72 hrs. In conclusion, these honeys and their extracts could be leads for further investigation in the discovery of new natural anti-H. pylori compounds.

Antimicrobial Potential and Phytochemical Screening of Clathria sp. 1 and Tedania (Tedania) stylonychaeta Sponge Crude Extracts Obtained from the South East Coast of South Africa.

Scientists have continuously searched for novel bioactive compounds to overcome the inherent problems associated with drug resistance, the evolution of unknown diseases, and the toxicity of currently used compounds. The ocean has been considered a rich source of compounds that possess unique chemical structures and novel biological capabilities. Biologically active molecules isolated from marine flora and fauna have shown significant advancement over the past century in the pharmaceutical industry. Marine natural products (MNPs) have been used as nanomedicine, cosmetics, wound healing, antimicrobial agents, anticancer agents, and anti-inflammatory agents. The physicochemical parameters of the collection site were also recorded. This study's marine sponge species were collected from Phillip's Reef, South Africa, at 12 m during the spring season. Ethyl acetate (EA) and dichloromethane : methanol (DCM : ME, 1 : 1) were used as extraction solvents. Crude extracts of the marine sponges were tested against MRSA, P. aeruginosa, C. difficile, A. fumigatus, and C. albicans. Phytochemical screening was conducted to identify seven critical phytochemical groups. A pH reading of 8.01 and a temperature of 15.45°C were recorded at the sampling site. Clathria sp. 1 and Tedania (Tedania) stylonychaeta EA crude extracts showed bioactivity against all five test pathogens. The DCM : ME crude extract of Clathria sp. 1 was the only bioactive crude extract from DCM : ME extracts. This crude extract was only bioactive against C. albicans as no activity was observed against the other four pathogens. EA crude extracts of Clathria sp. 1 yielded more significant inhibition zones against both fungal pathogens. These EA crude extracts performed better than fluconazole as inhibition zones of 35 ± 0 mm at 24 mg/ml, 31 ± 0 mm at 19 mg/ml, 31 ± 0 mm at 14.4 mg/ml, 30 ± 0 mm at 9.6 mg/ml, and 25 ± 0 mm at 7.2 mg/ml were recorded. Clathria sp. 1 crude extracts exhibited higher inhibition zones compared to Tedania (Tedania) stylonychaeta. The antibiotic imipenem (26 ± 0.7 mm at 10 µg) and ciprofloxacin (30 ± 0.3 mm at 5 µg) exhibited higher zones of inhibition than EA crude extracts of Tedania (Tedania) stylonychaeta at all test concentrations. In this study, Clathria sp. 1 was observed to have broad-spectrum bioactivity as EA crude extracts were bioactive against MRSA, P. aeruginosa, C. difficile, A. fumigatus, and C. albicans. In addition to this, the EA crude extract of Clathria sp. 1 was bacteriostatic (9.6 mg/ml). Clathria sp. 1 DCM : ME crude extract only tested positive for the presence of terpenoids. In contrast, EA crude extracts did not test positive for the existence of any of the seven phytochemicals. Our study has revealed that Tedania (Tedania) stylonychaeta and Clathria sp. 1 sponge species collected from Phillip's Reef in South Africa can produce bioactive compounds useful against bacterial and fungal species.

Seasonal Variation in Antimicrobial Activity of Crude Extracts of Psammaplysilla sp. 1 from Phillips Reef, South Africa.

Marine invertebrates constitute a diverse group of marine organisms beneficial to humanity due to their therapeutic significance. The marine sponge species Psammaplysilla sp. 1 was collected from Philips Reef, South Africa, over a four-season period and assayed for antimicrobial potential. The physicochemical parameters of the collection site were also recorded. The sponge crude extracts' antimicrobial activity was evaluated using an agar well diffusion assay against 5 pathogens. Phytochemical screening was conducted to identify the presence of 7 critical phytochemical groups. During the four seasons, the mean water temperature was 17.35°C ± 2.06, with autumn recording the highest (20°C) temperature. Antifungal activity was observed by Psammaplysilla sp. 1 (30 mm) against C. albicans, and this was higher than that showed by standard drugs ICZ-10 µg (15 ± 0.1 mm), FLU-15 µg (21 ± 0.2 mm), and VCZ-5 µg (17 ± 0.1 mm), respectively. Similar bioactivities were observed seasonally with Psammaplysilla sp. 1 (22 mm and 24 mm) during autumn and spring, respectively, against C. difficile while only crude extracts collected in spring showed bioactivity against C. albicans. Psammaplysilla sp. crude extracts showed broad-spectrum bioactivity against all test pathogens. DCM : ME crude extracts tested positive for the presence of 2/7 of the phytochemicals (terpenoids and flavonoids). GC-MS revealed several previously reported biologically active compounds such bicyclo[4.2.0]octa-1,3,5-trien-7-ol and phenol, 2,6-dibromo, some of which have been found in plants. This study revealed that sponge bioactivity is dependent on the season and further validated the antimicrobial potential of South African marine sponges.

Antibiotic Susceptibility of Non-Cholera Vibrios Isolated from Farmed and Wild Marine Fish (Argyrosomus japonicus), Implications for Public Health.

This study aimed to evaluate the antibiogram and antibiotic resistance genes (ARGs) of Vibrio isolates recovered from a marine fish (Argyrosomus japonicus) and water samples from two commercial dusky kob aquaculture farms and the Kariega estuary, South Africa, and to evaluate these findings for their public health implications. A total of 277 molecularly confirmed Vibrio isolates consisting of 126 Vibrio fluvialis, 45 Vibrio vulnificus, 30 Vibrio Parahaemolyticus, and 76 vibrios belonging to species of the genus other than Vibrio cholerae were subjected to susceptibility testing to 15 antibiotics by the disc diffusion method. Multiple antibiotic resistance index (MARI) was used to determine the antibiotic resistance-associated health risk, while polymerase chain reaction was used to evaluate the presence of 14 ARGs for nonsusceptible strains. Highest resistances were recorded to amoxicillin (76.2%), ampicillin (67.5%), erythromycin (38.3%), and doxycycline (35.0%), while susceptibilities were highest to gentamicin (100%), followed by norfloxacin (97.8%), florfenicol (90.3%), tetracycline (87.7%), and chloramphenicol (87.4%). We recorded a 58.5% multidrug resistance (resistance to ≥2 antimicrobial classes). MARI did not vary significantly between sites (p > 0.05); however, values of >0.2 were recorded in 40% (108/277) of all strains tested. ARG markers, ampC, blaOXA, tetA, tetM, dfr1, sul1, sul2, ermB, nptII, strA, and SXT integrase, were detected in one or more strains with ermB (82.5%), sul2 (53.8%), strA (44%), dfr1 (42.3%), and tetM (38.3%) being the most abundant. Healthy marine finfish (dusky kob) and their environment can serve as reservoirs for antibiotic resistant vibrios and ARGs, which could be disseminated to humans and other susceptible bacteria and this therefore becomes a public health concern.

The Effect of Polyherbal Medicines Used for the Treatment of Tuberculosis on Other Opportunistic Organisms of Humans Infected with Tuberculosis.

BACKGROUND: In many immunocompromised patients, opportunistic bacterial and fungal infections are common. Polyherbal medicines examined in this study are used by the indigenous people of South Africa for the treatment of tuberculosis (TB) and other opportunistic infections associated with TB. OBJECTIVE: To evaluate the antibacterial and antifungal activity of nine polyherbal remedies against four Gram-positive and Gram-negative bacteria respectively and three fungi. MATERIALS AND METHODS: Agar dilution method was used to determine the minimum inhibitory concentration (MIC) of the remedies against the organisms. RESULTS: The inhibitory activity of the polyherbal medicines based on the overall MIC revealed that HBfs and FB remedies were the most active remedies against the bacterial isolates at the concentration of 2.5 mg/mL, followed by HBts remedy at 5.0 mg/mL. However, the MIC valves of KWTa, KWTb, KWTc, HBss, EL and AL remedies were higher than 5.0 mg/mL which was the highest concentration used. Only KWTa remedy showed activity against Aspergillus niger and Aspergillus fumigatus with the MIC value of 2.5 mg/mL. While KWTc and HBts had the highest activity at 1.25 mg/mL against Candida albicans, the remaining remedies were active at 2.5 mg/mL. CONCLUSION: This study revealed that some of these polyherbal formulations have activities against some of the opportunistic bacterial and fungal isolates associated with TB patients. The capability of these remedies to inhibit the organisms is an indication that they are a potential broad-spectrum antimicrobial agent. However, the remedies that are inactive might contain stimulant effects on the immune system. SUMMARY: In the Eastern Cape Province of South Africa, no study has been reported on the effect of polyherbal remedies used for the treatment of TB on the opportunistic pathogen. This study therefore revealed that some of the polyherbal medicines possess activity against bacterial and fungal pathogens. Abbreviations used: TB: Tuberculosis; MIC: Minimum Inhibitory Concentration; CFU/ML: Colony Forming Unit Per Mill.

Occurrence of Virulence Genes Associated with Human Pathogenic Vibrios Isolated from Two Commercial Dusky Kob (Argyrosmus japonicus) Farms and Kareiga Estuary in the Eastern Cape Province, South Africa.

Background: Seafood-borne Vibrio infections, often linked to contaminated seafood and water, are of increasing global public health concern. The aim of this study was to evaluate the prevalence of human pathogenic vibrios and their associated virulence genes isolated from fish and water samples from 2 commercial dusky kob farms and Kareiga estuary, South Africa. Methods: A total of 200 samples including dusky kob fish (n = 120) and seawater (n = 80) were subjected to Vibrio screening on thiosulfate-citrate-bile salts-sucrose agar (TCBS). Presumptive isolates were confirmed and delineated to V. cholerae, V. parahaemolyticus, V. vulnificus, and V. fluvialis by PCR. Various pathogenic gene markers were screened: V. parahaemolyticus (trh and tdh), V. vulnificus (vcgE and vcgC) and V. fluvialis (stn, vfh,hupO, vfpA). Restriction Fragment Length Polymorphism (RFLP) of the vvhA gene of V. vulnificus strains was performed to determine the associated biotypes. Results: Total Vibrio prevalence was 59.4% (606/1020) of which V. fluvialis was the most predominant 193 (31.85%), followed by Vibrio vulnificus 74 (12.21%) and V. parahaemolyticus 33 (5.45%). No V. cholerae strain was detected. One of the V. parahaemolyticus strains possessed the trh gene 7 (9.46%) while most (91.9%; 68/74) V. vulnificus isolates were of the E-type genotype. V. fluvialis virulence genes detected were stn (13.5%), hupO (10.4%) and vfpA (1.0%). 12.16% (9/74) of V. vulnificus strains exhibited a biotype 3 RFLP pattern. Conclusions: This is the first report of potentially pathogenic vibrios from healthy marine fish in the study area, and therefore a public health concern.

Identification of bacterial contaminants in polyherbal medicines used for the treatment of tuberculosis in Amatole District of the Eastern Cape Province, South Africa, using rapid 16S rRNA technique.

BACKGROUND: Polyherbal medicines are used for the treatment of many diseases in many African and Asian communities. With the increasing use of these remedies, several investigations have shown that they are associated with a broad variety of residues and contaminants. This study investigates the presence of bacteria in the polyherbal medicines used for the treatment of tuberculosis (TB) in the Eastern Cape Province of South Africa. METHODS: Bacterial DNA was extracted from the polyherbal medicines, and a fragment of the bacterial 16S rRNA gene was amplified by PCR with universal primers 27F and 518R. The amplicons were visualised on agarose gel electrophoresis, followed by end repair and adaptor ligation. They were further purified and quantified using Library Preparation kit NEBNext® UltraT DNA Library Prep Kit for Illumina, and the amplicons were run on illumina's MiSeq platform. RESULTS: Different bacterial species were identified in all each of the polyherbal medicines. Generally, the most prominent and common bacteria recovered from all the samples were Bacillus sp., Enterobacter sp., Klebsiella sp., Rahnella sp., Paenibacillus sp., Clostridium sp. and Pantoea sp. Others are Pseudomonas sp., Raoultella ornithinolytica, Salmonella enterica and Eubacterium moniliforme. CONCLUSIONS: This study, thus, revealed the presence of pathogenic and non-pathogenic bacteria in the polyherbal medicines used for the treatment of tuberculosis in the study area. The implications of the findings are discussed in relation to the health care of the patients of tuberculosis in the study area, having in mind that they are immunocompromised individuals.