Investigation into the bacterial pollution in three Western Cape rivers, South Africa and the application of bioremediation strategies as clean-up technology

Abstract

The quality of South Africa’s water sources is fast deteriorating due to an influx of pollutants from industrial and agricultural areas. In addition, urbanisation has led to the establishment of informal settlements along river systems. This study focuses on the importance of maintaining water quality and the management of water resources in order to ensure its sustainability in South Africa. The primary aim of this study was to determine the extent of bacterial contamination in three rivers namely the Berg-, Plankenburg- and Diep Rivers in the Western Cape, South Africa and to investigate the application of a bioremediation system as a possible treatment technology. Several aspects contributing to the contamination were addressed and different approaches were studied and reviewed. In all three rivers, four sampling sites were identified, which were sampled over a period of 9 to 12 months. Contamination levels for the three rivers were evaluated by applying various enumeration techniques, which could provide an accurate indication of the planktonic bacterial pollution load in the river systems. The Most Probable Number (MPN) technique was used to determine the level of faecal coliforms and E. coli. The highest MPN, faecal coliform and E. coli counts of 3.5 x 107 micro-organisms/100 m , 3.5 x 107 micro-organisms/100 m and 1.7 x 107 micro-organisms/100 m , respectively, were recorded at Site B2 in week 37 in the Berg River. Results showed that in all the river water sampled and evaluated, the total MPN count mostly exceeded the maximum limit of 2000 micro-organisms/100 m (SABS, 1984) stipulated for river water throughout the study period. The heterotrophic plate count (HPC) method was used to determine the number of culturable micro-organisms in planktonic samples, while the flow cytometry (FCM) and epifluorescence microscopy (EM) with different fluorochromes (Acridine orange and BacLight™ Live/Dead stain) were employed to evaluate total bacterial counts in planktonic (water) samples. The highest HPC at the various sites sampled was 1.04 x 106 micro-organisms/m (Berg River, Site B2), 7.9 x 104 micro-organisms/m (Plankenbrug River, Site A) and 1.7 x 105 micro-organisms/m (Diep River, Site B). Total cell counts as high as 3.7 x 107 micro-organism/m (Berg River, Site B2), 5.5 x 108 micro-organism/m (Plankenburg River, Site D) and 2.5 x 109 micro-organisms/m (Diep River, Site B) were obtained by the FCM technique, which were significantly (p < 0.05) higher than the total counts obtained by epifluorescence microscopy. The results thus show that

the FCM technique was the most reliable method for determining the total cell count in river water samples. This technique makes use of computer software whereas epifluorescence microscopy involves manual counting which may lead to human error. In addition, the impact of residential, agricultural and industrial areas situated along these rivers was also investigated. Even though exact point sources of pollution could not be determined, it was found that all the sources, such as the storm water drainage pipes, the industrial as well as the agricultural areas, could contribute to increased MPN, heterotrophic and total bacterial counts. This study also aimed at investigating and comparing the microbial contamination levels at various sites in the Plankenburg and Diep Rivers in the Western Cape, South Africa. Sampling of sites along the Plankenburg River started in June 2004 and continued for a period of one year until June 2005. Sampling of the Diep River sites started in March 2005 and continued for a period of nine months until November 2005. Faecal coliform (FC) and E. coli (EC) counts were determined by means of the Most Probable Number technique, the number of culturable cells were determined using the heterotrophic plate count (HPC) technique and total microbial counts were evaluated by Flow cytometric analysis (FCM). The highest microbial counts for the Plankenburg River were observed at site B where the highest MPN, FC, E. coli and total FCM counts of 9.2 x 106 (week 14), 3.5 x 106 (week 39) and 3.5 x 106 micro-organisms/100 m (week 39) and 2.1 x 108 micro-organisms/m (weeks 1 and 39) respectively, were recorded. The highest HPC recorded for the Plankenburg River was 7.9 x 106 micro-organisms/100 m (week 44, site A). Site B is situated close to an informal settlement where waste effluents from storm water drainage pipes enter the river system. In addition, other possible contamination sources included agricultural (site A) and industrial (site C) areas bordering the Plankenburg River. The highest total MPN, FC and E. coli counts in the Diep River were 5.4 x 106 (week 23) and 1.6 x 106 micro-organisms/100 m [FC and E. coli, respectively (both in week 23)], recorded at site B. The highest HPC and total FCM counts of 1.7 x 107 micro-organisms/100 m (week 14) and 2.5 x 109 microorganisms/ m (week 23), respectively, were also recorded at site B. This site was identified as the most contaminated site along the Diep River and served as an accumulation point for waste effluents from the residential and industrial areas, which included paint and machine manufacturers. Other sources situated along the Diep River included storage and maintenance facilities for steel containers, a waste water treatment plant and an oil-refinery. Most of the bacterial counts obtained for the

Plankenburg and Diep Rivers exceeded the accepted maximum limit for river water for most of the sampling period. Bacterial species from the Berg- and Plankenburg Rivers were isolated and identified. The presence of various Enterobacteriaceae species isolated at all the sites in both rivers confirmed faecal contamination of these water sources over the entire sampling period. Opportunistic pathogens such as Klebsiella sp., Serratia sp., Enterobacter sp., Shewanella sp., Aeromonas sp., Pseudomonas sp., Acinetobacter sp. and Citrobacter freundii as well as pathogens such as Bacillus cereus and B. anthracis were also identified in both river systems. All the respective articles are presented in the required format of the journal in which the article has been published or submitted to.