Memory Tekere

Metagenomic analysis of bacterial diversity of Siloam hot water spring, Limpopo, South Africa

The bacterial diversity of Siloam hot water spring was determined using 454 pyrosequencing of two 16S rRNA variable regions V1-3 and V4-7. Analysis of the community DNA revealed that the phyla Proteobacteria, Cyanobacteria, Bacteriodetes, Planctomycetes, Firmicutes, Chloroflexi and Verrucomicrobia were the most abundant. The bacterial diversity detectable and classifiable was greater when the V4-7 variable region was used compared to the V1-3 region. The most abundant bacteria genera detected with region V1-3 were; Stenotrophomonas (23.3%), Aquaspirillum (5.11%) , Zavarzinella (2.73%), Haliscomenobacteria (1.25%), Rheinheimera (1.14%) and Tepidomonas (1.14%). All the other detectable genera were below 0.6%. Genera detected with region V4-7 from most abundant were; Stenotrophomonas (17.96%), Zavarzinella (5.81%), Aquaspirillum (4.75%), Rheinheimera (3.52%), GPI (1.41%), Gemmata (1.41%) and Syntrophobacter (1.06%). All the other genera detected were below 0.7%. Siloam is one of the hottest thermal springs in South Africa (63°C), the water has a pH of 9.5 and is relatively high in fluoride and bromide; it is possible that the physicochemical properties could have some influence on the diversity of bacteria. This article reports on the first phylogenetic analysis of a South African thermal spring bacterial community. Key words : Thermophilic, hot springs, biodiversity, 454 sequencing, South Africa.

Comparisons of the composition and biogeographic distribution of the bacterial communities occupying South African thermal springs with those inhabiting deep subsurface fracture water.

South Africa has numerous thermal springs that represent topographically driven meteoric water migrating along major fracture zones. The temperature (40–70°C) and pH (8–9) of the thermal springs in the Limpopo Province are very similar to those of the low salinity fracture water encountered in the South African mines at depths ranging from 1.0 to 3.1 km. The major cation and anion composition of these thermal springs are very similar to that of the deep fracture water with the exception of the dissolved inorganic carbon and dissolved O2, both of which are typically higher in the springs than in the deep fracture water. The in situ biological relatedness of such thermal springs and the subsurface fracture fluids that feed them has not previously been evaluated. In this study, we evaluated the microbial diversity of six thermal spring and six subsurface sites in South Africa using high-throughput sequencing of 16S rRNA gene hypervariable regions. Proteobacteria were identified as the dominant phylum within both subsurface and thermal spring environments, but only one genera, Rheinheimera, was identified among all samples. Using Morisita similarity indices as a metric for pairwise comparisons between sites, we found that the communities of thermal springs are highly distinct from subsurface datasets. Although the Limpopo thermal springs do not appear to provide a new window for viewing subsurface bacterial communities, we report that the taxonomic compositions of the subsurface sites studied are more similar than previous results would indicate and provide evidence that the microbial communities sampled at depth are more correlated to subsurface conditions than geographical distance.

Urban effluent discharges as causes of public and environmental health concerns in South Africa's aquatic milieu

The water quality in South Africa’s river systems is rapidly deteriorating as a consequence of increased discharge of wastewater effluents. The natural ability of rivers and reservoirs to trap toxic chemicals and nutrients in their sediments enables these systems to accumulate contaminants, altering the natural balance in environmental water quality, thereby raising a plethora of public and environmental health concerns. Impaired water quality has been linked to an array of problems in South Africa including massive fish mortalities, altered habitat template leading to the thinning of riverine macroinvertebrate diversity, shifts in microbial community structures with drastic ecological consequences and evolvement of antibiotic resistance genes that, under natural conditions, can be transferred to waterborne pathogens. Urban wastewater discharge has also been implicated in increased bioaccumulation of metals in edible plant parts, elevated concentrations of endocrine-disrupting compounds (EDCs), which are blamed for reduced fertility and increased cancer risk, excessive growth of toxic cyanobacteria and an increase in concentrations of pathogenic microorganisms which constitute a potential health threat to humans. However, despite the ecotoxicological hazards posed by wastewater effluents, ecotoxicological studies are currently underutilised in South African aquatic ecosystem assessments, and where they have been done, the observation is that ecotoxicological studies are mostly experimental and restricted to small study areas. More research is still needed to fully assess especially the ecotoxicological consequences of surface water pollution by urban wastewater effluents in South Africa. A review of the effects of urban effluent discharges that include domestic effluent mixed with industrial effluent and/or urban stormwater run-off is hereby presented.

An evaluation of the bacterial diversity at Tshipise, Mphephu and Sagole hot water springs, Limpopo Province, South Africa

This research was supported through a grant from Water Research Commission (WRC, SA, Project K5/1959/1).

Diversity Analysis and Bioresource Characterization of Halophilic Bacteria Isolated from a South African Saltpan

Though intensive research has been channeled towards the biotechnological applications of halophiles and other extremophilic microbes, these studies have not been, by any means, exhaustive. Saline environments still offer a vast diversity of microbes with potential to produce an array of natural products which can only be unlocked by concerted research efforts. In this study, a combination of culture and molecular approaches were employed to characterize halophilic bacteria from saltpan water samples and profile their potential biotechnological applications. Physicochemical analysis of the water samples showed that pH was alkaline (pH 8.8), with a salinity of 12.8%. 16S rRNA gene targeted amplicon analysis produced 10 bacterial phyla constituting of Bacteroidetes (30.57%), Proteobacteria (15.27%), Actinobacteria (9.05%), Planctomycetes (5.52%) and Cyanobacteria (3.18%). Eighteen strains were identified using sequencing analysis of the culturable bacterial strains. From these, the strains SP7 and SP9 were positive for cellulase production while the strains SP4, SP8 and SP22 were positive for lipase production. Quantitative enzyme assays showed moderate extracellular cellulase activity (1.95 U/mL) and lipase activity (3.71 U/mL) by the isolate SP9 and SP4 respectively. Further, of the six isolates, the isolate SP9 exhibited exploitable potential in the bioremediation of hydrocarbon pollution as demonstrated by its fairly high activity against benzanthracene (70% DCPIP reduction). Elucidation of the isolates secondary metabolites showed the production of the molecules 2,3-butanediol, hexahydro-3-(2-methylpropyl)pyrrole[1,2a]pyrazine-1,4-dione, aziridine, dimethylamine and ethyl acetate (GC-MS) and oxypurinol and 5-hydroxydecanoic acid (LC-MS), particularly by the isolate Salinivibrio sp. SP9. Overall, the study showed that the isolated halophiles can produce secondary metabolites with potential industrial and pharmaceutical application.

Evaluation of heavy metal removal from wastewater in a modified packed bed biofilm reactor

For the effective application of a modified packed bed biofilm reactor (PBBR) in wastewater industrial practice, it is essential to distinguish the tolerance of the system for heavy metals removal. The industrial contamination of wastewater from various sources (e.g. Zn, Cu, Cd and Ni) was studied to assess the impacts on a PBBR. This biological system was examined by evaluating the tolerance of different strengths of composite heavy metals at the optimum hydraulic retention time (HRT) of 2 hours. The heavy metal content of the wastewater outlet stream was then compared to the source material. Different biomass concentrations in the reactor were assessed. The results show that the system can efficiently treat 20 (mg/l) concentrations of combined heavy metals at an optimum HRT condition (2 hours), while above this strength there should be a substantially negative impact on treatment efficiency. Average organic reduction, in terms of the chemical oxygen demand (COD) of the system, is reduced above the tolerance limits for heavy metals as mentioned above. The PBBR biological system, in the presence of high surface area carrier media and a high microbial population to the tune of 10 000 (mg/l), is capable of removing the industrial contamination in wastewater.

Thermophilic bacterial communities inhabiting the microbial mats of “indifferent” and chalybeate (iron-rich) thermal springs: Diversity and biotechnological analysis

Microbial mats are occasionally reported in thermal springs and information on such mats is very scarce. In this study, microbial mats were collected from two hot springs (Brandvlei (BV) and Calitzdorp (CA)), South Africa and subjected to scanning electron microscopy (SEM) and targeted 16S rRNA gene amplicon analysis using Next Generation Sequencing (NGS). Spring water temperature was 55°C for Brandvlei and 58°C for Calitzdorp while the pH of both springs was slightly acidic, with an almost identical pH range (6.2–6.3). NGS analysis resulted in a total of 4943 reads, 517 and 736 OTUs for BV and CA at, respectively, a combined total of 14 different phyla in both samples, 88 genera in CA compared to 45 in BV and 37.64% unclassified sequences in CA compared to 27.32% recorded in BV. Dominant bacterial genera in CA microbial mat were Proteobacteria (29.19%), Bacteroidetes (9.41%), Firmicutes (9.01%), Cyanobacteria (6.89%), Actinobacteria (2.65%), Deinococcus‐Thermus (2.57%), and Planctomycetes (1.94%) while the BV microbial mat was dominated by Bacteroidetes (47.3%), Deinococcus‐Thermus (12.35%), Proteobacteria (7.98%), and Planctomycetes (2.97%). Scanning electron microscopy results showed the presence of microbial filaments possibly resembling cyanobacteria, coccids, rod‐shaped bacteria and diatoms in both microbial mats. Dominant genera that were detected in this study have been linked to different biotechnological applications including hydrocarbon degradation, glycerol fermentation, anoxic‐fermentation, dehalogenation, and biomining processes. Overall, the results of this study exhibited thermophilic bacterial community structures with high diversity in microbial mats, which have a potential for biotechnological exploitation.

Synthetic extreme environments: overlooked sources of potential biotechnologically relevant microorganisms

Synthetic extreme environments like carwash effluent tanks and drains are potential sources of biotechnologically important microorganisms and molecules which have, however, remained unexplored. Using culture‐ and molecular‐based methods, a total of 17 bacterial isolates belonging to the genera Shewanella, Proteus, Paenibacillus, Enterobacter and Citrobacter, Aeromonas, Pseudomonas and Pantoea were identified. Hydrocarbon utilization and enzyme production screening assays showed that Aeromonas sp. CAC11, Paenibacillus sp. CAC12 and Paenibacillus sp. CAC13 and Citrobacter sp. PCW7 were able to degrade benzanthracene, naphthalene and diesel oil, Paenibacillus sp. CAC12 and Paenibacillus sp. CAC13 could produce cellulase enzyme, while Proteus sp. BPS2, Pseudomonas sp. SAS8 and Proteus sp. CAL3 could produce lipase. GC‐MS analysis of bacterial secondary metabolites resulted in identification of 107 different compounds produced by Proteus sp. BPS2, Paenibacillus sp. CAC12, Pseudomonas sp. SAS8, Proteus sp. CAL3 and Paenibacillus sp. CAC13. Most of the compounds identified by both GC‐MS and LC‐MS have previously been determined to have antibacterial, antifungal and/or anticancer properties. Further, microbial metabolites which have previously been known to be produced only by plants or microorganisms found in natural extreme environments were also identified in this study. This research has revealed the immense bioresource potential of microorganisms inhabiting synthetic extreme environments.

Potential biotechnological capabilities of cultivable mycobiota from carwash effluents

Urban life has created man‐made extreme environments like carwashes. These environments have, however, not been sufficiently explored for mycobiota that can be sources of biotechnologically useful products, as has been the case with natural extreme environments. Using a combination of culture and molecular techniques, fungi from carwash effluents was characterized for production of lipase and cellulase enzymes, nonpolar and polar biotechnologically relevant secondary metabolites and hydrocarbon utilization. The isolated fungal strains belonged to the genera Alternaria, Cladosporium, Penicillium, Peyronellaea, Rhizopus, Spegazzinia, Trichoderma, Ulocladium and Yarrowia. Sixty‐six percent (66%) of the fungal isolates were found to be able to metabolize naphthalene and benzanthracene, showing potential for application in bioremediation of hydrocarbon polluted sites. Lipase production by the isolates Penicillium sp. BPS3 (2.61 U/ml), Trichoderma sp. BPS9 (2.01 U/ml), Rhizopus sp. CAL1 (2.05 U/ml), Penicillium sp. PCW1 (2.99 U/ml) and Penicillium sp. SAS1 (2.16 U/ml) compared well with previously recorded lipase production levels by other fungi. The highest producers of cellulase were Penicillium sp. SAS1 (12.10 U/ml), Peyronella sp. CAW5 (4.49 U/ml) and Cladosporium sp. SAS3 (4.07 U/ml), although these activities were lower than previously reported levels. GC‐MS analysis of the fungal secondary metabolites resulted in identification of 572 compounds, including azulene, methanamine, N‐pentylidene, metoclopramide, and mepivacaine while compounds determined by UHPLC‐MS included 10‐undecen‐1‐ol, piquerol A, 10‐undecyn‐1‐ol, cyclo(leucylprolyl) and rac‐etomidate. These compounds were previously determined to have various activities including anticancer, antibacterial, antifungal, antihypertensive, antidiabetic and anti‐inflammatory properties. The study demonstrated that fungi from carwash effluents are natural sources of some biotechnologically important products.