Assessment of the incidence of E.coli in Tyume and Buffalo rivers in the Eastern Cape Province of South Africa

• Main Author: Koba, Siziwe
• Format: Others
• Language: English
• Published: University of Fort Hare 2013
• Online Access: http://hdl.handle.net/10353/d1006889

Waterborne diseases are among the leading causes of morbidity and mortality in developing countries and every year around 2.2 million people die due to basic hygiene related diseases like coliform diarrhoea. Universal access to safe drinking water and sanitation has been promoted as an essential step in reducing these preventable diseases (Tambekar and Banginwar, 2005; Patil, 2004; Charan, 2004). Diarrheagenic Escherichia coli are one of the most important etiologic agents of acute diarrhea and represent a major public health problem in developing countries like South Africa The present study was conducted between August 2010 and July 2011 to investigate the prevalence and distribution of virulent E. coli strains from water samples collected from Tyume and Buffalo River, located in Eastern Cape Province of South Africa using conventional microbiological methods and PCR analysis. Twelve Water samples were collected from three different sites of the rivers, viz; upstream, middle stream and the downstream of the dam. E.coli was isolated by the membrane filtration method on mFC. A total of 374 isolates from both rivers were identified by convenctional microbiological techniques. For both Buffalo and Tyume River, A large proportion (87 and 114, respectively) of the isolates from the mid stream samples satisfied the identification characteristics for E. coli (blue colonies on MFC agar and violet/purple colonies on Chromocult agar) and thus revealing high levels contamination when compared to isolates from the downstream (55 and 47) and the upstream (30 and 31) All the isolates that satisfied the primary identification stage were subjected to PCR. DNA was extracted using the boiling method and then the DNA was used as a template for PCR. Specific PCR analysis was performed on all E. coli isolates to amplify the alr gene that codes for alanine racemase Out and of the 202 isolates amplified for Tyume river, 70 (35 percent) were positively identified as E. coli since they possessed the alr gene fragment. and out of the 172 isolates amplified from Buffalo River, 80(47 percent) were also positively identified as E. coli. For both Tyume and Buffalo River, the highest prevalence was observed midstream (39 percent and 56 pecent respectively). The identified E. coli were further characterized into different pathotypes. Amplification of the shig gene, LT gene, EaeA gene, Eagg gene and the ST gene were used to detect pathogenic E.coli. In Tyume River, Genes of ETEC (lt or st) were detected in 21/70 (30 percent); Gene of EPEC (eae) was detected in 14/70 specimens (35 percent); Genes of EAEC (Eagg) was detected in 14/70(35 percent) and genes of EIEC (shig) were detected in 11/70(16 percent). In Buffalo River, no DEC was recovered upstream and downstream of the river. EAEC (8 percent) was the only pathotypes recovered midstream of the river. Strains of all five E. coli categories showed high-level resistance to ampicillin, tetracycline, cotrimoxazole, and chloramphenicol but were highly susceptible to quinolones, aminoglycosides, and novobiocin. The highest resistance (100 percent) amongst the isolates was observed to ampicillin by EAEC, Heat Labile (ETEC) and EIEC, followed by 87.5 percent by EAEC to carbenicillin. The highest susceptibility was to quinolones (100 percent) by all the four categories of E.coli. The screening for antibiotic resistance genes revealed the absence of SHV, CTMX and TetC genes as they were not detected in any of the E.coli isolates. However, TEM genes were observed in 80 percent of the isolates. Integron conserved segment was detected in these same organisms in the same proportion as TEM