Assessment of the quality indices and prevalence of Escherichia coli pathotypes in selected rivers of Osun state, Southwestern Nigeria
- Authors: Titilawo, Osuolale Yinka
- Date: 2015
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11295 , http://hdl.handle.net/10353/d1021274
- Description: Surface waters are important freshwater sources used for domestic, industrial, agricultural and recreational activities, and the availability of good quality freshwater is indispensable for preventing water-borne diseases and improving quality of life especially in communities that lack pipe-borne water. Water samples were collected from ten rivers at different locations in Osun State, Southwestern Nigeria. A total of 12 physicochemical parameters, counts of total coliforms (TC) and Escherichia coli isolates were determined using standard analytical procedures. Confirmed Escherichia coli isolates (n=300) were assessed for the presence of 10 virulence genes (VGs) associated with Escherichia coli strains causing intestinal and extra-intestinal infections. The recovered Escherichia coli isolates were elucidated for their antibiogram profiling by disk diffusion method and the resistant isolates were further profiled for their genotypic antimicrobial resistance by polymerase chain reaction technique. The physicochemical qualities ranged as follows: pH (6.9 - 7.6), temperature (26 – 29 ºC), turbidity (2.28 – 9.46 NTU), electrical conductivity (229 – 581 μS/cm), nitrate (0.03 – 0.05 mg/L), nitrite (0.00 – 0.01 mg/L), sulphate (3.33 – 20.33 mg/L), chloride ions (7.83 – 27.33 mg/L), dissolved oxygen (4.23 – 5.57 mg/L), total dissolved solids (56 – 184 mg/L), total hardness (78 – 519 mg/L) and alkalinity (50.67 – 146.67 mg/L). Statistical analysis showed that pH, temperature, electrical conductivities, nitrates, nitrites, chloride, dissolved oxygen, total dissolved solid, total hardness and alkalinity were significantly different (P < 0.05), whereas turbidity and sulphate were not significantly different (P ˃ 0.05) from each parameter with respect to sampling sites. While the VG lt for enterotoxigenic E. coli had the highest prevalence of 45%, the enteropathogenic E. coli genes eae and bfp were detected in 6% and 4% of the isolates respectively. The VGs stx1 and stx2 specific for the enterohemorrhagic E. coli pathotypes were equally detected in 7% and 1% of the isolates respectively. Also, the VG eagg harboured by enteroaggregative pathotype and diffusely-adherent E. coli VG daaE were detected in 2% and 4% of the isolates respectively and enteroinvasive E. coli VG ipaH was not detected. In addition, the VGs papC for uropathogenic and ibeA for neonatal meningitis were frequently detected in 19% and 3% of isolates respectively. While all the isolates tested were susceptible to imipenem, meropenem, amikacin and gatilofloxacin, others were variously susceptible, and resistant as follows; ciprofloxacin (96%), kanamycin (95%), neomycin (92%), streptomycin (84%), chloramphenicol (73%), nalidixic acid (66%), nitrofuratoin (64%), gentamycin (63%), doxycycline (58%), cefepime (57%), tetracycline (49%) and cephalothin (42%). Conversely, all the isolates were resistant to sulphamethoxazole, and high levels of resistance were equally observed against amoxycillin (59%), ampicilin (57%) and cefuroxime (40%). Cefepime, cephalothin, cefuroxime, nalidixic acid, nitrofuratoin, chloramphenicol and tetracycline were not significantly different in their effect against the isolates from all locations (P > 0.05), whereas the resistance profile of the isolates against gentamycin, ciprofloxacin, sulphamethoxazole, ampicillin and amoxicillin were significantly different (P < 0.05). Amikacin, kanamycin, streptomycin, meropenem, imipenem and gatilofloxacin were statistically excluded from the analysis since all tested isolates showed total susceptibility to these antimicrobials. The multiple antibiotic resistance indexing ranged from 0.50 to 0.80 for all the sampling locations and exceeded the threshold value of 0.2. Prevalence and distributions of the 19 resistance determinants assessed were obtained as follows; [sulfonamides (sulI (8%), sulII (41%)], [beta-lactams; (ampC 22%; blaTEM, (21%), blaZ (18%),], [tetracyclines (tetA (24%), tetB (23%), tetC (18%), tetD (78%), tetK (15%), tetM, (10%)], [phenicols; (catI (37%), catII (28%), cmlA1 (19%)] and [aminoglycosides; (aacC2 (8%), aphA1 (80%), aphA2 (80%), aadA (79%) and strA (38%)]. The Pearson chi square exact test revealed many strong significant associations among ampC, blaTEM, blaZ and tetA genes with some determinants screened. In the same vein, a grand total of 366 resistance gene fingerprints were spotted across the sampling locations and among the resistant pathotypes, the modal prevalent gene prints were found among the ETEC strains in 148 (40%), being the predominant pathotype observed, followed by UPEC strains 80 (22%) while the lowest was the least occurring EAEC pathotype 14 (4%). While some physicochemical parameters exceeded prescribed standards for drinking water, some fell within. The total coliforms obtained in all the sampling sites were above the acceptable limits. Findings reveal the presence of diarrhoeagenic and non-diarrhoeagenic E. coli in the selected rivers and suggest a potential public health risk as the rivers are important resources for domestic, recreational and livelihood usage by their host communities. The multiple drug resistance indexing signifies isolates and pathotypes of high antimicrobial usage origin. An increase in the antimicrobial resistance signatures towards conventionally used antibiotics as observed in this study necessitates for safe water supply, adequate sanitation facilities and proper surveillance programs towards the monitoring of antimicrobial resistance determinants in water-bodies. Generally, results from this study indicate that the river waters are not suitable for consumption, domestic or recreational use and re-echo the importance of safeguarding the freshwater resources of Southwestern Nigeria.
- Full Text:
- Date Issued: 2015
- Authors: Titilawo, Osuolale Yinka
- Date: 2015
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11295 , http://hdl.handle.net/10353/d1021274
- Description: Surface waters are important freshwater sources used for domestic, industrial, agricultural and recreational activities, and the availability of good quality freshwater is indispensable for preventing water-borne diseases and improving quality of life especially in communities that lack pipe-borne water. Water samples were collected from ten rivers at different locations in Osun State, Southwestern Nigeria. A total of 12 physicochemical parameters, counts of total coliforms (TC) and Escherichia coli isolates were determined using standard analytical procedures. Confirmed Escherichia coli isolates (n=300) were assessed for the presence of 10 virulence genes (VGs) associated with Escherichia coli strains causing intestinal and extra-intestinal infections. The recovered Escherichia coli isolates were elucidated for their antibiogram profiling by disk diffusion method and the resistant isolates were further profiled for their genotypic antimicrobial resistance by polymerase chain reaction technique. The physicochemical qualities ranged as follows: pH (6.9 - 7.6), temperature (26 – 29 ºC), turbidity (2.28 – 9.46 NTU), electrical conductivity (229 – 581 μS/cm), nitrate (0.03 – 0.05 mg/L), nitrite (0.00 – 0.01 mg/L), sulphate (3.33 – 20.33 mg/L), chloride ions (7.83 – 27.33 mg/L), dissolved oxygen (4.23 – 5.57 mg/L), total dissolved solids (56 – 184 mg/L), total hardness (78 – 519 mg/L) and alkalinity (50.67 – 146.67 mg/L). Statistical analysis showed that pH, temperature, electrical conductivities, nitrates, nitrites, chloride, dissolved oxygen, total dissolved solid, total hardness and alkalinity were significantly different (P < 0.05), whereas turbidity and sulphate were not significantly different (P ˃ 0.05) from each parameter with respect to sampling sites. While the VG lt for enterotoxigenic E. coli had the highest prevalence of 45%, the enteropathogenic E. coli genes eae and bfp were detected in 6% and 4% of the isolates respectively. The VGs stx1 and stx2 specific for the enterohemorrhagic E. coli pathotypes were equally detected in 7% and 1% of the isolates respectively. Also, the VG eagg harboured by enteroaggregative pathotype and diffusely-adherent E. coli VG daaE were detected in 2% and 4% of the isolates respectively and enteroinvasive E. coli VG ipaH was not detected. In addition, the VGs papC for uropathogenic and ibeA for neonatal meningitis were frequently detected in 19% and 3% of isolates respectively. While all the isolates tested were susceptible to imipenem, meropenem, amikacin and gatilofloxacin, others were variously susceptible, and resistant as follows; ciprofloxacin (96%), kanamycin (95%), neomycin (92%), streptomycin (84%), chloramphenicol (73%), nalidixic acid (66%), nitrofuratoin (64%), gentamycin (63%), doxycycline (58%), cefepime (57%), tetracycline (49%) and cephalothin (42%). Conversely, all the isolates were resistant to sulphamethoxazole, and high levels of resistance were equally observed against amoxycillin (59%), ampicilin (57%) and cefuroxime (40%). Cefepime, cephalothin, cefuroxime, nalidixic acid, nitrofuratoin, chloramphenicol and tetracycline were not significantly different in their effect against the isolates from all locations (P > 0.05), whereas the resistance profile of the isolates against gentamycin, ciprofloxacin, sulphamethoxazole, ampicillin and amoxicillin were significantly different (P < 0.05). Amikacin, kanamycin, streptomycin, meropenem, imipenem and gatilofloxacin were statistically excluded from the analysis since all tested isolates showed total susceptibility to these antimicrobials. The multiple antibiotic resistance indexing ranged from 0.50 to 0.80 for all the sampling locations and exceeded the threshold value of 0.2. Prevalence and distributions of the 19 resistance determinants assessed were obtained as follows; [sulfonamides (sulI (8%), sulII (41%)], [beta-lactams; (ampC 22%; blaTEM, (21%), blaZ (18%),], [tetracyclines (tetA (24%), tetB (23%), tetC (18%), tetD (78%), tetK (15%), tetM, (10%)], [phenicols; (catI (37%), catII (28%), cmlA1 (19%)] and [aminoglycosides; (aacC2 (8%), aphA1 (80%), aphA2 (80%), aadA (79%) and strA (38%)]. The Pearson chi square exact test revealed many strong significant associations among ampC, blaTEM, blaZ and tetA genes with some determinants screened. In the same vein, a grand total of 366 resistance gene fingerprints were spotted across the sampling locations and among the resistant pathotypes, the modal prevalent gene prints were found among the ETEC strains in 148 (40%), being the predominant pathotype observed, followed by UPEC strains 80 (22%) while the lowest was the least occurring EAEC pathotype 14 (4%). While some physicochemical parameters exceeded prescribed standards for drinking water, some fell within. The total coliforms obtained in all the sampling sites were above the acceptable limits. Findings reveal the presence of diarrhoeagenic and non-diarrhoeagenic E. coli in the selected rivers and suggest a potential public health risk as the rivers are important resources for domestic, recreational and livelihood usage by their host communities. The multiple drug resistance indexing signifies isolates and pathotypes of high antimicrobial usage origin. An increase in the antimicrobial resistance signatures towards conventionally used antibiotics as observed in this study necessitates for safe water supply, adequate sanitation facilities and proper surveillance programs towards the monitoring of antimicrobial resistance determinants in water-bodies. Generally, results from this study indicate that the river waters are not suitable for consumption, domestic or recreational use and re-echo the importance of safeguarding the freshwater resources of Southwestern Nigeria.
- Full Text:
- Date Issued: 2015
Evaluation of the quality indices of the final effluents of two wastewater treatment plants in buffalo City Metropolitan Municipality in the Eastern Cape Province
- Authors: Osuolale, Olayinka O
- Date: 2015
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11292 , http://hdl.handle.net/10353/d1021079
- Description: Waste waters can be sources of pollution to surface water and the environment with severe implications for public health. Most treatment plants in the Buffalo City Municipality in the Eastern Cape Province discharge their treated effluent into the surface waters which directly and indirectly impacts on the quality of surface waters in the region. The objective of this study was to determine the microbiological and physicochemical qualities of the final effluents of two wastewater treatment plants in the Buffalo City Municipality in the Eastern Cape Province of South Africa over a period of 12 months (September 2012 to August 2013). The qualities of the final effluents of WW-Ama Wastewater Treatment Plant with respect to phosphate (3.9 mg/l - 20.6 mg/l), free chlorine (0.05 mg/l - 0.71 mg/l), chemical oxygen demand (COD) (4.7 mg/l - 211 mg/l), and faecal coliform (0 - 2.92 × 104 CFU/100 ml) were not in compliance with the permissible limits set for effluent discharged to surface water by South Africa guidelines for effluent discharge. Other physicochemical parameters like biological oxygen demand (BOD) (2.2 mg/l - 9.0 mg/l), total dissolve solid (TDS) (253 mg/l - 336.3 mg/l) and turbidity (4.8 NTU - 43.20 NTU) with no SA regulatory set limits were compared to other regulatory standards and they do not comply with the limits. Also, at the second WWTP’s, the WW-Dim Treatment Plant effluent quality for free chlorine (0.06 mg/l - 7.2 mg/l), BOD (0.1 mg/l - 7.4 mg/l), and turbidity (4.02 NTU - 24.3 NTU) also did not comply. For microbiological qualities, counts of presumptive E. coli and Vibrio ranged between 0 - 2.92 × PROFESSOR ANTHONY I. OKOHAntibiogram of the bacterial isolates were determined using the disk diffusion method. A total of 107 confirmed E. coli and 100 confirmed Vibrio spp. were used for this assay. Results of antibiotic sensitivity test revealed that 63.6% of the E. coli isolates were resistance to ampicillin while 49.5% were resistant to tetracycline and cephalothin. The least resistances were observed against gentamicin (3.7%) and cefotaxime (1.9%). No resistance was observed against meropenem. For the Vibrio spp, resistance was most frequently observed against tetracycline (38%) ampicillin (26%), chloramphenicol (16%), cefotaxime (14%), trimethoprim-sulfamethoxazole (13%) and the least resistance observed was against ciprofloxacin (1%). This study demonstrates that poorly treated wastewater effluent can be a source of eutrophic water with high nutrient levels and pathogenic bacteria and enteric viruses as well as antibiotic resistance determinants that could impact negatively on human health. The finding of this study also suggests that WWTPs have to be properly monitored and controlled to ensure compliance to set guidelines. This could be attained through the application of appropriate treatment processes, which will help to minimize possible dangers to public environment health.
- Full Text:
- Date Issued: 2015
- Authors: Osuolale, Olayinka O
- Date: 2015
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11292 , http://hdl.handle.net/10353/d1021079
- Description: Waste waters can be sources of pollution to surface water and the environment with severe implications for public health. Most treatment plants in the Buffalo City Municipality in the Eastern Cape Province discharge their treated effluent into the surface waters which directly and indirectly impacts on the quality of surface waters in the region. The objective of this study was to determine the microbiological and physicochemical qualities of the final effluents of two wastewater treatment plants in the Buffalo City Municipality in the Eastern Cape Province of South Africa over a period of 12 months (September 2012 to August 2013). The qualities of the final effluents of WW-Ama Wastewater Treatment Plant with respect to phosphate (3.9 mg/l - 20.6 mg/l), free chlorine (0.05 mg/l - 0.71 mg/l), chemical oxygen demand (COD) (4.7 mg/l - 211 mg/l), and faecal coliform (0 - 2.92 × 104 CFU/100 ml) were not in compliance with the permissible limits set for effluent discharged to surface water by South Africa guidelines for effluent discharge. Other physicochemical parameters like biological oxygen demand (BOD) (2.2 mg/l - 9.0 mg/l), total dissolve solid (TDS) (253 mg/l - 336.3 mg/l) and turbidity (4.8 NTU - 43.20 NTU) with no SA regulatory set limits were compared to other regulatory standards and they do not comply with the limits. Also, at the second WWTP’s, the WW-Dim Treatment Plant effluent quality for free chlorine (0.06 mg/l - 7.2 mg/l), BOD (0.1 mg/l - 7.4 mg/l), and turbidity (4.02 NTU - 24.3 NTU) also did not comply. For microbiological qualities, counts of presumptive E. coli and Vibrio ranged between 0 - 2.92 × PROFESSOR ANTHONY I. OKOHAntibiogram of the bacterial isolates were determined using the disk diffusion method. A total of 107 confirmed E. coli and 100 confirmed Vibrio spp. were used for this assay. Results of antibiotic sensitivity test revealed that 63.6% of the E. coli isolates were resistance to ampicillin while 49.5% were resistant to tetracycline and cephalothin. The least resistances were observed against gentamicin (3.7%) and cefotaxime (1.9%). No resistance was observed against meropenem. For the Vibrio spp, resistance was most frequently observed against tetracycline (38%) ampicillin (26%), chloramphenicol (16%), cefotaxime (14%), trimethoprim-sulfamethoxazole (13%) and the least resistance observed was against ciprofloxacin (1%). This study demonstrates that poorly treated wastewater effluent can be a source of eutrophic water with high nutrient levels and pathogenic bacteria and enteric viruses as well as antibiotic resistance determinants that could impact negatively on human health. The finding of this study also suggests that WWTPs have to be properly monitored and controlled to ensure compliance to set guidelines. This could be attained through the application of appropriate treatment processes, which will help to minimize possible dangers to public environment health.
- Full Text:
- Date Issued: 2015
Assessment of the anti-Listerial properties of Garcinia kola (Heckel) seeds
- Authors: Penduka, Dambudzo
- Date: 2014
- Subjects: Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11278 , http://hdl.handle.net/10353/d1015527 , Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Description: A follow-up of traditional medicinal plants uses is an important tool in highlighting their therapeutic potentials, as they have been found to be a source of a wide range of bioactive compounds that can be used as base compounds for new pharmaceutical drugs. This study therefore focuses on assessing the anti-Listerial properties of the seeds of Garcinia kola (Heckel) plant, which is a traditional medicinal plant of west and central African origin, and was and is still used to traditionally treat several ailments. Four different solvents crude extracts of the seeds were assessed for their anti-Listerial activities in-vitro, against a panel of 42 Listeria bacteria, which included Listeria monocytogenes, Listeria ivanovii and Listeria grayi species. At 10 mg/ml concentration the aqueous extract had activity against 29% of the test isolates while the other three crude extracts namely dichloromethane, n-hexane and the methanol extracts had activity against 45% of the test bacteria. The minimum inhibitory concentration (MIC) ranges of the extracts were 0.079-0.313 mg/ml for the dichloromethane extract; 0.079-0.625 mg/ml for the n-hexane extract; 0.157-0.625 mg/ml for the methanol extract; and 10->10 mg/ml for the aqueous extract. The minimum bactericidal concentration (MBC) ranges of the extracts were 0.625–10 mg/ml for both the n-hexane and the dichloromethane extract; 5-10 mg/ml for the methanol extract; and those for the aqueous extract were above 10 mg/ml against all the susceptible Listeria isolates. The rate of kill analysis was then determined for the three most active crude extracts that is excluding the aqueous extract and it was assessed against four representative Listeria species namely L. monocytogenes (LAL 8), L. grayi (LAL 15), L. ivanovii (LEL 30) and L. ivanovii (LEL 18). All the three extracts showed a general trend of being concentration and time dependent in their rate of kill profiles such that most bacteria cells were killed at the highest test concentration of 4× MIC value after the maximum exposure time of 2 h. The n-hexane, dichloromethane and methanol extracts were bactericidal against 4, 3 and 1 isolates out of the four test Listeria isolates respectively.
- Full Text:
- Date Issued: 2014
- Authors: Penduka, Dambudzo
- Date: 2014
- Subjects: Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11278 , http://hdl.handle.net/10353/d1015527 , Microbial sensitivity tests , Garcinia , Medicinal plants , Traditional medicine
- Description: A follow-up of traditional medicinal plants uses is an important tool in highlighting their therapeutic potentials, as they have been found to be a source of a wide range of bioactive compounds that can be used as base compounds for new pharmaceutical drugs. This study therefore focuses on assessing the anti-Listerial properties of the seeds of Garcinia kola (Heckel) plant, which is a traditional medicinal plant of west and central African origin, and was and is still used to traditionally treat several ailments. Four different solvents crude extracts of the seeds were assessed for their anti-Listerial activities in-vitro, against a panel of 42 Listeria bacteria, which included Listeria monocytogenes, Listeria ivanovii and Listeria grayi species. At 10 mg/ml concentration the aqueous extract had activity against 29% of the test isolates while the other three crude extracts namely dichloromethane, n-hexane and the methanol extracts had activity against 45% of the test bacteria. The minimum inhibitory concentration (MIC) ranges of the extracts were 0.079-0.313 mg/ml for the dichloromethane extract; 0.079-0.625 mg/ml for the n-hexane extract; 0.157-0.625 mg/ml for the methanol extract; and 10->10 mg/ml for the aqueous extract. The minimum bactericidal concentration (MBC) ranges of the extracts were 0.625–10 mg/ml for both the n-hexane and the dichloromethane extract; 5-10 mg/ml for the methanol extract; and those for the aqueous extract were above 10 mg/ml against all the susceptible Listeria isolates. The rate of kill analysis was then determined for the three most active crude extracts that is excluding the aqueous extract and it was assessed against four representative Listeria species namely L. monocytogenes (LAL 8), L. grayi (LAL 15), L. ivanovii (LEL 30) and L. ivanovii (LEL 18). All the three extracts showed a general trend of being concentration and time dependent in their rate of kill profiles such that most bacteria cells were killed at the highest test concentration of 4× MIC value after the maximum exposure time of 2 h. The n-hexane, dichloromethane and methanol extracts were bactericidal against 4, 3 and 1 isolates out of the four test Listeria isolates respectively.
- Full Text:
- Date Issued: 2014
Assessment of the incidence of E.coli in Tyume and Buffalo rivers in the Eastern Cape Province of South Africa
- Authors: Koba, Siziwe
- Date: 2013
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11272 , http://hdl.handle.net/10353/d1006889
- Description: 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
- Full Text:
- Date Issued: 2013
- Authors: Koba, Siziwe
- Date: 2013
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11272 , http://hdl.handle.net/10353/d1006889
- Description: 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
- Full Text:
- Date Issued: 2013
Antibacterial and phytochemical studies of selected South African honeys on clinical isolates of Helicobacter pylori
- Authors: Manyi-Loh, Christy E
- Date: 2012
- Subjects: Helicobacter pylori , Honey--South Africa , Drug resistance in microorganisms , Bacterial diseases , Honey -- Therapeutic use , Helicobacter pylori infections , Traditional medicine -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11240 , http://hdl.handle.net/10353/d1001056 , Helicobacter pylori , Honey--South Africa , Drug resistance in microorganisms , Bacterial diseases , Honey -- Therapeutic use , Helicobacter pylori infections , Traditional medicine -- South Africa -- Eastern Cape
- Description: Infection with Helicobacter pylori has been associated with the pathogenesis of numerous stomach and gastroduodenal diseases that pose threats to public health. Eradicaftion of this pathogen is a global challenge due to its alarming rate of multidrug resistance. Consequently, to find an alternative treatment, the search is increasingly focused on new antimicrobial product from natural sources including honey. Honey has been used as medicine in several cultures since ancient time due to its enormous biomedical activities. Its beneficial qualities have been endorsed to its antimicrobial, antioxidant, anti-inflammatory properties added to its phytocomponents. In this study, the anti-H. pylori activity of South African honeys and their solvent extracts as well as the phytochemicals present in the two most active honeys were evaluated. Agar well diffusion test was used to investigate the antimicrobial activity of six honey varieties obtained from different locations in the country. Subsequently, the honeys were extracted with four organic solvents viz n-hexane, diethyl ether, chloroform and ethyl acetate employed in order of increasing polarity. The antibacterial activity of the different solvent extracts of each honey was evaluated by agar well diffusion; broth micro dilution and time kill assays. Different chromatographic techniques (Thin layer & column chromatography) were employed to enumerate the phytochemical constituents in the most active solvent extracts of Pure Honey (PH) and Champagne Royal Train (CRT); and were identified by gas-chromatography linked mass-spectrometry. Linalool pure compound was equally evaluated for anti-H. pylori activity in a bid to trace the antibacterial agent among the variety of compounds identified. Data were analyzed by One-way ANOVA test at 95% confidence interval. Crude honeys and their solvent extracts demonstrated potent anti-H. pylori activity with zone diameter that ranged from [16.0mm (crude) to 22.2mm (extract)] and percentage susceptibilities of test isolates between 73.3% (crude) and 93.3% (extract). The chloroform extracts of PH and CRT were most active with MIC50 in the ranges 0.01- viii 10%v/v and 0.625-10%v/v respectively, not significantly different from amoxicillin (P> 0.05); and efficient bactericidal activity (100% bacterial cells killed) at 1/2MIC and 4xMIC over different time intervals, 36-72hrs and 18-72hrs respectively. The appearance of bands on the thin layer chromatography (TLC) chromatogram spotted with the chloroform extracts of PH and CRT; and developed with hexane: ethyl acetate: acetic acid (HEA) and methanol: acetic acid: water (MAAW) solvent systems indicated the presence of compounds. Purification of the compounds contained in these extracts over silica gel column yielded numerous fractions which were evaluated for antibacterial activity and purity. PHF5 was the most active fraction with a mean MIC50 value of 1.25mg/mL. Volatile compounds belonging to different known chemical families in honey were identified in all the active fractions obtained from PH. Conversely, only four compounds were identified in the active fractions obtained from CRT hence the non volatile constituents could be of prime relevance with respect to antibacterial activity of this honey. Of novelty was the presence of thiophene and N-methyl-D3-azirdine compounds, essential precursors used for the synthesis of natural products and pharmaceuticals with vital biomedical properties. Linalool demonstrated potent inhibitory (MIC95, 0.002- 0.0313mg/mL) and bactericidal activity (0.0039-0.313mg/mL) against the test isolates. On the other hand, a significant difference was recorded (P < 0.05) in comparing the activity of linalool compound to the fractions. PH could serve as a good economic source of bioactive compounds which could be employed as template for the synthesis of novel anti-H. pylori drugs. However, further studies are needed to determine the non volatile active ingredients in PH and CRT as well as toxicological testing
- Full Text:
- Date Issued: 2012
- Authors: Manyi-Loh, Christy E
- Date: 2012
- Subjects: Helicobacter pylori , Honey--South Africa , Drug resistance in microorganisms , Bacterial diseases , Honey -- Therapeutic use , Helicobacter pylori infections , Traditional medicine -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11240 , http://hdl.handle.net/10353/d1001056 , Helicobacter pylori , Honey--South Africa , Drug resistance in microorganisms , Bacterial diseases , Honey -- Therapeutic use , Helicobacter pylori infections , Traditional medicine -- South Africa -- Eastern Cape
- Description: Infection with Helicobacter pylori has been associated with the pathogenesis of numerous stomach and gastroduodenal diseases that pose threats to public health. Eradicaftion of this pathogen is a global challenge due to its alarming rate of multidrug resistance. Consequently, to find an alternative treatment, the search is increasingly focused on new antimicrobial product from natural sources including honey. Honey has been used as medicine in several cultures since ancient time due to its enormous biomedical activities. Its beneficial qualities have been endorsed to its antimicrobial, antioxidant, anti-inflammatory properties added to its phytocomponents. In this study, the anti-H. pylori activity of South African honeys and their solvent extracts as well as the phytochemicals present in the two most active honeys were evaluated. Agar well diffusion test was used to investigate the antimicrobial activity of six honey varieties obtained from different locations in the country. Subsequently, the honeys were extracted with four organic solvents viz n-hexane, diethyl ether, chloroform and ethyl acetate employed in order of increasing polarity. The antibacterial activity of the different solvent extracts of each honey was evaluated by agar well diffusion; broth micro dilution and time kill assays. Different chromatographic techniques (Thin layer & column chromatography) were employed to enumerate the phytochemical constituents in the most active solvent extracts of Pure Honey (PH) and Champagne Royal Train (CRT); and were identified by gas-chromatography linked mass-spectrometry. Linalool pure compound was equally evaluated for anti-H. pylori activity in a bid to trace the antibacterial agent among the variety of compounds identified. Data were analyzed by One-way ANOVA test at 95% confidence interval. Crude honeys and their solvent extracts demonstrated potent anti-H. pylori activity with zone diameter that ranged from [16.0mm (crude) to 22.2mm (extract)] and percentage susceptibilities of test isolates between 73.3% (crude) and 93.3% (extract). The chloroform extracts of PH and CRT were most active with MIC50 in the ranges 0.01- viii 10%v/v and 0.625-10%v/v respectively, not significantly different from amoxicillin (P> 0.05); and efficient bactericidal activity (100% bacterial cells killed) at 1/2MIC and 4xMIC over different time intervals, 36-72hrs and 18-72hrs respectively. The appearance of bands on the thin layer chromatography (TLC) chromatogram spotted with the chloroform extracts of PH and CRT; and developed with hexane: ethyl acetate: acetic acid (HEA) and methanol: acetic acid: water (MAAW) solvent systems indicated the presence of compounds. Purification of the compounds contained in these extracts over silica gel column yielded numerous fractions which were evaluated for antibacterial activity and purity. PHF5 was the most active fraction with a mean MIC50 value of 1.25mg/mL. Volatile compounds belonging to different known chemical families in honey were identified in all the active fractions obtained from PH. Conversely, only four compounds were identified in the active fractions obtained from CRT hence the non volatile constituents could be of prime relevance with respect to antibacterial activity of this honey. Of novelty was the presence of thiophene and N-methyl-D3-azirdine compounds, essential precursors used for the synthesis of natural products and pharmaceuticals with vital biomedical properties. Linalool demonstrated potent inhibitory (MIC95, 0.002- 0.0313mg/mL) and bactericidal activity (0.0039-0.313mg/mL) against the test isolates. On the other hand, a significant difference was recorded (P < 0.05) in comparing the activity of linalool compound to the fractions. PH could serve as a good economic source of bioactive compounds which could be employed as template for the synthesis of novel anti-H. pylori drugs. However, further studies are needed to determine the non volatile active ingredients in PH and CRT as well as toxicological testing
- Full Text:
- Date Issued: 2012
Phytochemical analysis and bioactivity of selected South African medicinal plants on clinical isolates of Helicobacter pylori
- Authors: Njume, Collise
- Date: 2011
- Subjects: Helicobacter pylori , Medicinal plants -- Biotechnology , Traditional medicine -- South Africa -- Eastern Cape , Antibiotics , Drug resistance in microorganisms , Extracts , Helicobacter pylori infections
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11260 , http://hdl.handle.net/10353/449 , Helicobacter pylori , Medicinal plants -- Biotechnology , Traditional medicine -- South Africa -- Eastern Cape , Antibiotics , Drug resistance in microorganisms , Extracts , Helicobacter pylori infections
- Description: Medicinal plants have been used as traditional medicine in the treatment of numerous human diseases for thousands of years in many parts of the world. In the developing world, especially in rural areas, herbal remedies continue to be a primary source of medicine. Scientifically, medicinal plants have proven to be an abundant source of biologically active compounds, many of which have already been formulated into useful therapeutic substances or have provided a basis for the development of new lead molecules for pharmaceuticals. Antibiotic resistance, undesireable side effects and expences associated with the use of combination therapy in the treatment of Helicobacter pylori infections have generated a considerable interest in the study of medicinal plants as potential sources of new drugs against this organism. The high complexicity of bioactive compounds accumulated in plants coupled with their broad antimicrobial activity may make it difficult for pathogenic organisms, including H. pylori to acquire resistance during treatment. This study therefore evaluates the antimicrobial potential of selected South African medicinal plants employed in the treatment of H. pylori-related infections, and the subsequent isolation of the plant active principles. An ethnobotanical survey of plants used in the treatment of H. pylori-related infections was conducted in the study area. Crude extracts of Combretum molle, Sclerocarya birrea, Garcinia kola, Alepidea amatymbica and 2 Strychnos species were screened against 30 clinical strains of H. pylori and 2 standard control strains (NCTC 11638 and ATCC 43526). In the preliminary stages of this study, ethyl acetate, acetone, ethanol, methanol and water extracts of the plants were tested against H. pylori by agar well diffusion and micro broth dilution methods. The plant crude extracts that exhibited anti-H. pylori activity with a iv percentage susceptibility of 50 percent and above were considered for the rate of kill assays and the most active crude extracts selected for bio-assay guided isolation of the active ingredient. Preliminary fractionation of the crude extract was achieved by thin layer chromatography (TLC) using different solvent combinations; hexane/diethylether (HDE), ethyl acetate/methanol/water (EMW) and chloroform/ethyl acetate/formic acid (CEF) in order to determine the most suitable combination for column chromatography (CC) and subsequent testing by indirect bioautography. The extract was then fractionated in a silica gel column using previously determined solvent combinations as eluent. Active fractions obtained from column chromatography separations were further fractionated and the compounds identified by gas chromatography/mass spectrometry (GC/MS) analysis. All the plants exhibited antimicrobial activity against H. pylori with zone of inhibition diameters ranging from 0 - 38 mm and minimum inhibitory concentration (MIC) values ranging from 0.06 - 5.0 mg/mL. The most active plant extracts were the acetone extract of C. molle with a percentage susceptibility of 87.1 percent, acetone and aqueous extracts of S. birrea (71 percent each) and the ethanolic extracts of G. kola (53.3 percent). Except for the aqueous extract, these extracts also exhibited a strong bactericidal activity against H. pylori at different concentrations. TLC analysis revealed the presence of 9 components in the acetone extract of S. birrea with the EMW solvent system as opposed to 5 and 8 with HDE and CEF respectively. Bioassay-guided isolation led to the identification of 52 compounds from the acetone extract of S. birrea with n-octacosane being the most abundant (41.68 percent). This was followed by pyrrolidine (38.91 percent), terpinen-4-ol (38.3 percent), n-eicosane (24.98 percent), cyclopentane (16.76 percent), n-triacontane (16.28 percent), aromadendrene (13.63 percent) and α-gujunene (8.77 percent). Terpinen-4-ol and pyrrolidine demonstrated strong antimicrobial activity against H. pylori at all concentrations tested. These results may serve as preliminary scientific validation of the ethnomedicinal uses of the above mentioned plants in the treatment of H. pylori-related infections in South Africa. Terpinen-4-ol and pyrrolidine could be considered for further evaluation as therapeutic or prophylactic agents in the treatment of H. pylori-related infections. However, further investigations would be necessary to determine their toxicological properties, in-vivo potencies and mechanism of action against H.pylori
- Full Text:
- Date Issued: 2011
- Authors: Njume, Collise
- Date: 2011
- Subjects: Helicobacter pylori , Medicinal plants -- Biotechnology , Traditional medicine -- South Africa -- Eastern Cape , Antibiotics , Drug resistance in microorganisms , Extracts , Helicobacter pylori infections
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11260 , http://hdl.handle.net/10353/449 , Helicobacter pylori , Medicinal plants -- Biotechnology , Traditional medicine -- South Africa -- Eastern Cape , Antibiotics , Drug resistance in microorganisms , Extracts , Helicobacter pylori infections
- Description: Medicinal plants have been used as traditional medicine in the treatment of numerous human diseases for thousands of years in many parts of the world. In the developing world, especially in rural areas, herbal remedies continue to be a primary source of medicine. Scientifically, medicinal plants have proven to be an abundant source of biologically active compounds, many of which have already been formulated into useful therapeutic substances or have provided a basis for the development of new lead molecules for pharmaceuticals. Antibiotic resistance, undesireable side effects and expences associated with the use of combination therapy in the treatment of Helicobacter pylori infections have generated a considerable interest in the study of medicinal plants as potential sources of new drugs against this organism. The high complexicity of bioactive compounds accumulated in plants coupled with their broad antimicrobial activity may make it difficult for pathogenic organisms, including H. pylori to acquire resistance during treatment. This study therefore evaluates the antimicrobial potential of selected South African medicinal plants employed in the treatment of H. pylori-related infections, and the subsequent isolation of the plant active principles. An ethnobotanical survey of plants used in the treatment of H. pylori-related infections was conducted in the study area. Crude extracts of Combretum molle, Sclerocarya birrea, Garcinia kola, Alepidea amatymbica and 2 Strychnos species were screened against 30 clinical strains of H. pylori and 2 standard control strains (NCTC 11638 and ATCC 43526). In the preliminary stages of this study, ethyl acetate, acetone, ethanol, methanol and water extracts of the plants were tested against H. pylori by agar well diffusion and micro broth dilution methods. The plant crude extracts that exhibited anti-H. pylori activity with a iv percentage susceptibility of 50 percent and above were considered for the rate of kill assays and the most active crude extracts selected for bio-assay guided isolation of the active ingredient. Preliminary fractionation of the crude extract was achieved by thin layer chromatography (TLC) using different solvent combinations; hexane/diethylether (HDE), ethyl acetate/methanol/water (EMW) and chloroform/ethyl acetate/formic acid (CEF) in order to determine the most suitable combination for column chromatography (CC) and subsequent testing by indirect bioautography. The extract was then fractionated in a silica gel column using previously determined solvent combinations as eluent. Active fractions obtained from column chromatography separations were further fractionated and the compounds identified by gas chromatography/mass spectrometry (GC/MS) analysis. All the plants exhibited antimicrobial activity against H. pylori with zone of inhibition diameters ranging from 0 - 38 mm and minimum inhibitory concentration (MIC) values ranging from 0.06 - 5.0 mg/mL. The most active plant extracts were the acetone extract of C. molle with a percentage susceptibility of 87.1 percent, acetone and aqueous extracts of S. birrea (71 percent each) and the ethanolic extracts of G. kola (53.3 percent). Except for the aqueous extract, these extracts also exhibited a strong bactericidal activity against H. pylori at different concentrations. TLC analysis revealed the presence of 9 components in the acetone extract of S. birrea with the EMW solvent system as opposed to 5 and 8 with HDE and CEF respectively. Bioassay-guided isolation led to the identification of 52 compounds from the acetone extract of S. birrea with n-octacosane being the most abundant (41.68 percent). This was followed by pyrrolidine (38.91 percent), terpinen-4-ol (38.3 percent), n-eicosane (24.98 percent), cyclopentane (16.76 percent), n-triacontane (16.28 percent), aromadendrene (13.63 percent) and α-gujunene (8.77 percent). Terpinen-4-ol and pyrrolidine demonstrated strong antimicrobial activity against H. pylori at all concentrations tested. These results may serve as preliminary scientific validation of the ethnomedicinal uses of the above mentioned plants in the treatment of H. pylori-related infections in South Africa. Terpinen-4-ol and pyrrolidine could be considered for further evaluation as therapeutic or prophylactic agents in the treatment of H. pylori-related infections. However, further investigations would be necessary to determine their toxicological properties, in-vivo potencies and mechanism of action against H.pylori
- Full Text:
- Date Issued: 2011
Assessment of the prevalence of virulent Eschericia coli strains in the final effluents of wastewater treatment plants in the Eastern Cape Province of South Africa
- Authors: Osode, Augustina Nwabuje
- Date: 2010
- Subjects: Escherichia coli , Escherichia coli -- Genetics , Effluent quality -- Testing , Water -- Purification , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Whole effluent toxicity testing
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11246 , http://hdl.handle.net/10353/d1001062 , Escherichia coli , Escherichia coli -- Genetics , Effluent quality -- Testing , Water -- Purification , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Whole effluent toxicity testing
- Description: Escherichia coli (E. coli) is a common inhabitant of surface waters in the developed and developing worlds. The majority of E. coli cells present in water are not particularly pathogenic to humans; however, there are some present in small proportion that possess virulence genes that allow them to colonize the digestive tract. Pathogenic E. coli causes acute and chronic diarrheal diseases, especially among children in developing countries and in travelers in these locales. The present study, conducted between August 2007 and July 2008, investigated the prevalence and distribution of virulent E. coli strains as either free or attached cells in the final effluents of three wastewater treatment plants located in the Eastern Cape Province of South Africa and its impact on the physico-chemical quality of the receiving water body. The wastewater treatment plants are located in urban (East Bank Reclamation Works, East London), peri-urban (Dimbaza Sewage Treatment Works) and in rural area (Alice Sewage Treatment Works). The effluent quality of the treatment plants were acceptable with respect to pH (6.9-7.8), temperature (13.8-22.0 °C), dissolved oxygen (DO) (4.9-7.8 mg/L), salinity (0.12-0.17 psu), total dissolved solids (TDS) (119-162 mg/ L) and nitrite concentration (0.1-0.4 mg/l). The other xii physicochemical parameters that did not comply with regulated standards include the following: phosphate (0.1-4.0 mg/L); chemical oxygen demand (COD) (5-211 mg/L); electrical conductivity (EC) (237-325 μS/cm) and Turbidity (7.7-62.7 NTU). Results suggest that eutrophication is intensified in the vicinity of the effluent discharge points, where phosphate and nitrate were found in high concentrations. Presumptive E. coli was isolated from the effluent samples by culture-based methods and confirmed using Polymerase Chain Reaction (PCR) techniques. Antibiogram assay was also carried out using standard in vitro methods on Mueller Hinton agar. The viable counts of presumptive E. coli for the effluent samples associated with 180 μm plankton size ranged between 0 – 4.30 × 101 cfu/ml in Dimbaza, 0 – 3.88 × 101 cfu/ml in Alice and 0 – 8.00 × 101 cfu/ml in East London. In the 60 μm plankton size category E. coli densities ranged between 0 and 4.2 × 101 cfu/ml in Dimbaza, 0 and 2.13 × 101 cfu/ml in Alice and 0 and 8.75 × 101 cfu/ml in East London. Whereas in the 20 μm plankton size category presumptive E. coli density varied from 0 to 5.0 × 101 cfu/ml in Dimbaza, 0 to 3.75 × 101 cfu/ml in Alice and 0 to 9.0 × 101 cfu/ml in East London. The free-living presumptive E. coli density ranged between 0 and 3.13 × 101 cfu/ml in Dimbaza, between 0 and 8.0 × 101 cfu/ml in Alice and between 0 and 9.5 × 101 cfu/ml in East London. Molecular analysis successfully amplified target genes (fliCH7, rfbEO157, ial and aap) which are characteristic of pathogenic E. coli strains. The PCR assays using uidA-specific primer confirmed that a genetic region homologous in size to the E. coli uidA structural gene, including the regulatory region, was present in 3 of the E. coli isolates from Alice, 10 from Dimbaza and 8 from East London. Of the 3 E. coli isolates from Alice, 1 (33.3%) was positive for the fliCH7 genes and 3 was positive for rfbEO157 genes. Out of the 10 isolates from Dimbaza, 4 were xiii positive for fliCH7 genes, 6 were positive for the rfbEO157 genes and 1 was positive for the aap genes; and of the 8 isolates from East London, 1 was positive for fliCH7 genes, 2 were for the rfbEO157 genes, 6 were positive for the ial genes. Antimicrobial susceptibility profile revealed that all of the E. coli strains isolated from the effluent water samples were resistant (R) to linezolid, polymyxin B, penicillin G and sulfamethoxazole. The E. coli isolates from Dimbaza (9/10) and East London (8/8) respectively were resistant to erythromycin. All the isolates were found to be susceptible (S) to amikacin, ceftazidime, ciprofloxacin, colistin sulphate, ceftriaxone, cefotaxime, cefuroxime, ertapenem, gatifloxacin, gentamycin, imidazole, kanamycin, meropenem, moxifloxacin, neomycin, netilmicin, norfloxacin and tobramycin. The findings of this study revealed that the Alice wastewater treatment plant was the most efficient as it produced the final effluent with the least pathogenic E. coli followed by the Dimbaza wastewater treatment plant. In addition, the findings showed that the wastewater treatment plant effluents are a veritable source of pathogenic E. coli in the Eastern Cape Province watershed. We suggest that to maximize public health protection, treated wastewater effluent quality should be diligently monitored pursuant to ensuring high quality of final effluents.
- Full Text:
- Date Issued: 2010
- Authors: Osode, Augustina Nwabuje
- Date: 2010
- Subjects: Escherichia coli , Escherichia coli -- Genetics , Effluent quality -- Testing , Water -- Purification , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Whole effluent toxicity testing
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11246 , http://hdl.handle.net/10353/d1001062 , Escherichia coli , Escherichia coli -- Genetics , Effluent quality -- Testing , Water -- Purification , Sewage disposal plants , Escherichia coli -- South Africa -- Eastern Cape , Whole effluent toxicity testing
- Description: Escherichia coli (E. coli) is a common inhabitant of surface waters in the developed and developing worlds. The majority of E. coli cells present in water are not particularly pathogenic to humans; however, there are some present in small proportion that possess virulence genes that allow them to colonize the digestive tract. Pathogenic E. coli causes acute and chronic diarrheal diseases, especially among children in developing countries and in travelers in these locales. The present study, conducted between August 2007 and July 2008, investigated the prevalence and distribution of virulent E. coli strains as either free or attached cells in the final effluents of three wastewater treatment plants located in the Eastern Cape Province of South Africa and its impact on the physico-chemical quality of the receiving water body. The wastewater treatment plants are located in urban (East Bank Reclamation Works, East London), peri-urban (Dimbaza Sewage Treatment Works) and in rural area (Alice Sewage Treatment Works). The effluent quality of the treatment plants were acceptable with respect to pH (6.9-7.8), temperature (13.8-22.0 °C), dissolved oxygen (DO) (4.9-7.8 mg/L), salinity (0.12-0.17 psu), total dissolved solids (TDS) (119-162 mg/ L) and nitrite concentration (0.1-0.4 mg/l). The other xii physicochemical parameters that did not comply with regulated standards include the following: phosphate (0.1-4.0 mg/L); chemical oxygen demand (COD) (5-211 mg/L); electrical conductivity (EC) (237-325 μS/cm) and Turbidity (7.7-62.7 NTU). Results suggest that eutrophication is intensified in the vicinity of the effluent discharge points, where phosphate and nitrate were found in high concentrations. Presumptive E. coli was isolated from the effluent samples by culture-based methods and confirmed using Polymerase Chain Reaction (PCR) techniques. Antibiogram assay was also carried out using standard in vitro methods on Mueller Hinton agar. The viable counts of presumptive E. coli for the effluent samples associated with 180 μm plankton size ranged between 0 – 4.30 × 101 cfu/ml in Dimbaza, 0 – 3.88 × 101 cfu/ml in Alice and 0 – 8.00 × 101 cfu/ml in East London. In the 60 μm plankton size category E. coli densities ranged between 0 and 4.2 × 101 cfu/ml in Dimbaza, 0 and 2.13 × 101 cfu/ml in Alice and 0 and 8.75 × 101 cfu/ml in East London. Whereas in the 20 μm plankton size category presumptive E. coli density varied from 0 to 5.0 × 101 cfu/ml in Dimbaza, 0 to 3.75 × 101 cfu/ml in Alice and 0 to 9.0 × 101 cfu/ml in East London. The free-living presumptive E. coli density ranged between 0 and 3.13 × 101 cfu/ml in Dimbaza, between 0 and 8.0 × 101 cfu/ml in Alice and between 0 and 9.5 × 101 cfu/ml in East London. Molecular analysis successfully amplified target genes (fliCH7, rfbEO157, ial and aap) which are characteristic of pathogenic E. coli strains. The PCR assays using uidA-specific primer confirmed that a genetic region homologous in size to the E. coli uidA structural gene, including the regulatory region, was present in 3 of the E. coli isolates from Alice, 10 from Dimbaza and 8 from East London. Of the 3 E. coli isolates from Alice, 1 (33.3%) was positive for the fliCH7 genes and 3 was positive for rfbEO157 genes. Out of the 10 isolates from Dimbaza, 4 were xiii positive for fliCH7 genes, 6 were positive for the rfbEO157 genes and 1 was positive for the aap genes; and of the 8 isolates from East London, 1 was positive for fliCH7 genes, 2 were for the rfbEO157 genes, 6 were positive for the ial genes. Antimicrobial susceptibility profile revealed that all of the E. coli strains isolated from the effluent water samples were resistant (R) to linezolid, polymyxin B, penicillin G and sulfamethoxazole. The E. coli isolates from Dimbaza (9/10) and East London (8/8) respectively were resistant to erythromycin. All the isolates were found to be susceptible (S) to amikacin, ceftazidime, ciprofloxacin, colistin sulphate, ceftriaxone, cefotaxime, cefuroxime, ertapenem, gatifloxacin, gentamycin, imidazole, kanamycin, meropenem, moxifloxacin, neomycin, netilmicin, norfloxacin and tobramycin. The findings of this study revealed that the Alice wastewater treatment plant was the most efficient as it produced the final effluent with the least pathogenic E. coli followed by the Dimbaza wastewater treatment plant. In addition, the findings showed that the wastewater treatment plant effluents are a veritable source of pathogenic E. coli in the Eastern Cape Province watershed. We suggest that to maximize public health protection, treated wastewater effluent quality should be diligently monitored pursuant to ensuring high quality of final effluents.
- Full Text:
- Date Issued: 2010
Prevalence of listeria pathogens in effluents of some wastewater treatment facilities in the Eastern Cape province of South Africa
- Odjadjare, Emmanuel Erufuare Onogwuwhenya
- Authors: Odjadjare, Emmanuel Erufuare Onogwuwhenya
- Date: 2010
- Subjects: Listeria -- South Africa -- Eastern Cape , Sewage -- Purification -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11264 , http://hdl.handle.net/10353/246 , Listeria -- South Africa -- Eastern Cape , Sewage -- Purification -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Description: Wastewater discharges may contain health compromising pathogens and carcinogenic and/or chemical substances that could compromise the public health and impact negatively on the environment. The present study was conducted between August 2007 and July 2008 to evaluate the Listeria abundance (as free-living and plankton associated species) and physicochemical qualities of the final effluents of three wastewater treatment facilities in the Eastern Cape Province of South Africa selected to represent typical urban, peri-urban and rural communities and the impact of the discharged final effluents on their respective receiving watershed, as well as to elucidated the in vitro antibiotic susceptibilities and resistance genes profile of Listeria species isolated from the final effluents. The suitability of the secondary effluent of the urban treatment facility (as a case study) for use in agriculture and aquaculture with reference to recommended standards was also determined. Wastewater samples were collected from the raw sewage, secondary effluent, final treated effluent, discharge point, 500 m upstream discharge point, and 500 m downstream discharge point from all three locations on a monthly basis throughout the study period. Listeria abundance in the final effluents and the receiving watersheds varied between 2.9× 100 and 3.52 × 105cfu/ml across the sampled locations. Free-living listerial density across the sampled locations ranged between 0 and 3.2 × 103cfu/ml while counts of Listeria species attached to large (180 μm) planktons varied from 0 to 1.58 × 105 cfu/ml and those of the 60 and 20 μm categories were in the range of 0 to 1.32 × 103 cfu/ml and 0 to 2.82 × 105 cfu/ml respectively. Listeria abundance did not vary significantly with location and season; there was however, significant (P < 0.05; P < 0.01) variance in Listeria abundance with plankton sizes across the locations. Free-living Listeria species were more abundant in the rural and urban xii communities than plankton attached Listeria species; whereas the reverse was the case in the peri-urban community. Prevalence of Listeria in terms of total counts was 100 percent across all sampled locations. Free-living Listeria species showed prevalence ranging from 84-96 percent across the sampling locations; while Listeria species attached to large (180 μm) planktons exhibited prevalence ranging from 75 percent to 90 percent. The prevalence of medium-sized (60 μm) plankton associated Listeria species varied between 58 percent and 92.5 percent; whereas those of Listeria species attached to small (20 μm) planktons ranged from 65-100 percent across all three communities. Listeria prevalence was generally a reflection of the turbidity of the water system, with free-living Listeria species being more prevalent than plankton associated cells in the relatively less turbid rural and urban waters compared to the more turbid peri-urban waters where plankton attached cells were more prevalent in comparison with their free living counterparts The final treated effluent quality fell short of recommended standards for turbidity, chemical oxygen demand and phosphate across all three communities. In addition, the final effluent of the rural treatment plant also fell short of recommended standard for NO3, while that of the urban treatment plant did not comply with acceptable limits for dissolved oxygen and nitrite. Other physicochemical parameters were compliant with set standards after treatment. An inverse relationship was observed between chlorine residual and listerial density across the sampled facilities; the effect of chlorine was however not enough to eliminate the pathogen from the water systems. At the urban treatment plant and its receiving watershed, pH, temperature, EC, turbidity, TDS, DO, and nitrate varied significantly with season and sampling point (P < 0.05; P < 0.01). Salinity also varied significantly with sampling point (P < 0.01), while COD and nitrite varied significantly with season (P < 0.05). Although, the treated effluent fell within recommended water quality standard for pH, TDS, nitrate and nitrite, it fell short of stipulated standards for other parameters. Whereas the microbial quality of the secondary treated effluent at this (urban) facility fell short of recommended standard after secondary treatment, its physicochemical quality were generally compliant with recommended standards for reuse wastewater in agriculture and aquaculture. Listeria pathogens isolated from effluents of the rural wastewater facility were sensitive to 11 (55 percent) of the 20 test antibiotics, and showed varying (7-71 percent) levels of resistance to 8 antibiotics; whereas those isolated from the peri-urban community showed sensitivity to 6 (30 percent) of the 20 test antibiotics, and varying (6-94 percent) levels of resistance to 12 antibiotics; while the urban effluent isolates were sensitive to 3 (15 percent) of the 20 test antibiotics, and showed varying (4.5-91 percent) levels of resistance to 17 antibiotics. Multiple antibiotic resistances involving 78.5-100 percent of isolates and antibiotics combination ranging from 2-10 antibiotics was observed across the sampled locations. Penicillin G and ampicillin showed remarkably high (64-91 percent) phenotypic resistance across the three sampled facilities. Other antibiotics, to which isolates showed significant resistance, were linezolid (22-88 percent); erythromycin (43-94 percent) and sulphamethoxazole (7-94 percent). Two of the 14 Listeria strains isolated from the rural effluents were positive for ereA and sul1 antibiotic resistance genes; while sulII genes were detected in five of the 23 Listeria isolates from the urban effluent and none was detected in isolates from the peri-urban community. The presence of antimicrobial resistance genes in the isolates did not correlate with phenotypic antibiotic resistance. The current study demonstrated that Listeria pathogens easily survived the activated sludge treatment process as free-living and plankton attached entities and suggests that municipal wastewater treatment plants are a significant source of multiple resistant Listeria pathogens in the South African aquatic milieu. While the physicochemical quality of the urban final effluent suggests that it is a major source of pollution to the receiving watershed, the secondary effluent quality demonstrated a great potential for use in agriculture and aquaculture.
- Full Text:
- Date Issued: 2010
- Authors: Odjadjare, Emmanuel Erufuare Onogwuwhenya
- Date: 2010
- Subjects: Listeria -- South Africa -- Eastern Cape , Sewage -- Purification -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11264 , http://hdl.handle.net/10353/246 , Listeria -- South Africa -- Eastern Cape , Sewage -- Purification -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Description: Wastewater discharges may contain health compromising pathogens and carcinogenic and/or chemical substances that could compromise the public health and impact negatively on the environment. The present study was conducted between August 2007 and July 2008 to evaluate the Listeria abundance (as free-living and plankton associated species) and physicochemical qualities of the final effluents of three wastewater treatment facilities in the Eastern Cape Province of South Africa selected to represent typical urban, peri-urban and rural communities and the impact of the discharged final effluents on their respective receiving watershed, as well as to elucidated the in vitro antibiotic susceptibilities and resistance genes profile of Listeria species isolated from the final effluents. The suitability of the secondary effluent of the urban treatment facility (as a case study) for use in agriculture and aquaculture with reference to recommended standards was also determined. Wastewater samples were collected from the raw sewage, secondary effluent, final treated effluent, discharge point, 500 m upstream discharge point, and 500 m downstream discharge point from all three locations on a monthly basis throughout the study period. Listeria abundance in the final effluents and the receiving watersheds varied between 2.9× 100 and 3.52 × 105cfu/ml across the sampled locations. Free-living listerial density across the sampled locations ranged between 0 and 3.2 × 103cfu/ml while counts of Listeria species attached to large (180 μm) planktons varied from 0 to 1.58 × 105 cfu/ml and those of the 60 and 20 μm categories were in the range of 0 to 1.32 × 103 cfu/ml and 0 to 2.82 × 105 cfu/ml respectively. Listeria abundance did not vary significantly with location and season; there was however, significant (P < 0.05; P < 0.01) variance in Listeria abundance with plankton sizes across the locations. Free-living Listeria species were more abundant in the rural and urban xii communities than plankton attached Listeria species; whereas the reverse was the case in the peri-urban community. Prevalence of Listeria in terms of total counts was 100 percent across all sampled locations. Free-living Listeria species showed prevalence ranging from 84-96 percent across the sampling locations; while Listeria species attached to large (180 μm) planktons exhibited prevalence ranging from 75 percent to 90 percent. The prevalence of medium-sized (60 μm) plankton associated Listeria species varied between 58 percent and 92.5 percent; whereas those of Listeria species attached to small (20 μm) planktons ranged from 65-100 percent across all three communities. Listeria prevalence was generally a reflection of the turbidity of the water system, with free-living Listeria species being more prevalent than plankton associated cells in the relatively less turbid rural and urban waters compared to the more turbid peri-urban waters where plankton attached cells were more prevalent in comparison with their free living counterparts The final treated effluent quality fell short of recommended standards for turbidity, chemical oxygen demand and phosphate across all three communities. In addition, the final effluent of the rural treatment plant also fell short of recommended standard for NO3, while that of the urban treatment plant did not comply with acceptable limits for dissolved oxygen and nitrite. Other physicochemical parameters were compliant with set standards after treatment. An inverse relationship was observed between chlorine residual and listerial density across the sampled facilities; the effect of chlorine was however not enough to eliminate the pathogen from the water systems. At the urban treatment plant and its receiving watershed, pH, temperature, EC, turbidity, TDS, DO, and nitrate varied significantly with season and sampling point (P < 0.05; P < 0.01). Salinity also varied significantly with sampling point (P < 0.01), while COD and nitrite varied significantly with season (P < 0.05). Although, the treated effluent fell within recommended water quality standard for pH, TDS, nitrate and nitrite, it fell short of stipulated standards for other parameters. Whereas the microbial quality of the secondary treated effluent at this (urban) facility fell short of recommended standard after secondary treatment, its physicochemical quality were generally compliant with recommended standards for reuse wastewater in agriculture and aquaculture. Listeria pathogens isolated from effluents of the rural wastewater facility were sensitive to 11 (55 percent) of the 20 test antibiotics, and showed varying (7-71 percent) levels of resistance to 8 antibiotics; whereas those isolated from the peri-urban community showed sensitivity to 6 (30 percent) of the 20 test antibiotics, and varying (6-94 percent) levels of resistance to 12 antibiotics; while the urban effluent isolates were sensitive to 3 (15 percent) of the 20 test antibiotics, and showed varying (4.5-91 percent) levels of resistance to 17 antibiotics. Multiple antibiotic resistances involving 78.5-100 percent of isolates and antibiotics combination ranging from 2-10 antibiotics was observed across the sampled locations. Penicillin G and ampicillin showed remarkably high (64-91 percent) phenotypic resistance across the three sampled facilities. Other antibiotics, to which isolates showed significant resistance, were linezolid (22-88 percent); erythromycin (43-94 percent) and sulphamethoxazole (7-94 percent). Two of the 14 Listeria strains isolated from the rural effluents were positive for ereA and sul1 antibiotic resistance genes; while sulII genes were detected in five of the 23 Listeria isolates from the urban effluent and none was detected in isolates from the peri-urban community. The presence of antimicrobial resistance genes in the isolates did not correlate with phenotypic antibiotic resistance. The current study demonstrated that Listeria pathogens easily survived the activated sludge treatment process as free-living and plankton attached entities and suggests that municipal wastewater treatment plants are a significant source of multiple resistant Listeria pathogens in the South African aquatic milieu. While the physicochemical quality of the urban final effluent suggests that it is a major source of pollution to the receiving watershed, the secondary effluent quality demonstrated a great potential for use in agriculture and aquaculture.
- Full Text:
- Date Issued: 2010
Surveillance of invasive vibro species in discharged aqueous efflents of wastewater treatment plants in the Eastern Cape province of South Africa
- Authors: Igbinosa, Etinosa Ogbomoede
- Date: 2010
- Subjects: Vibrio -- South Africa -- Eastern Cape , Water -- Fluoridation -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11267 , http://hdl.handle.net/10353/245 , Vibrio -- South Africa -- Eastern Cape , Water -- Fluoridation -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Description: Vibrio infections remain a serious threat to public health. In the last decade, Vibrio disease outbreaks have created a painful awareness of the personal, economic, societal, and public health costs associated with the impact of contaminated water in the aquatic milieu. This study was therefore designed to assess the prevalence of Vibrio pathogens in the final effluents of wastewater treatment plants (WWTPs) in the Eastern Cape Province, as well as their abilities to survive the treatment processes of the activated sludge system either as free cells or as plankton-associated entities in relation to the physicochemical qualities of the effluents. Three wastewater treatment facilities were selected to represent typical urban, sub-urban and rural communities, and samples were collected monthly from August 2007 to July 2008 from the final effluent, discharge point, 500 meter upstream and downstream of the discharge points and analysed for physicochemical parameters, Vibrio pathogens prevalence and their antibiogram characteristics using both culture based and molecular techniques. Physicochemical parameters measured include pH, temperature, electrical conductivity, salinity, turbidity, total dissolved solid (TDS), dissolved oxygen (DO), chemical oxygen demand (COD), nitrate, nitrite and orthophosphate levels. Unacceptably high levels of the assayed parameters were observed in many cases for COD (<10 - 1180 mg/l), nitrate (0.08 - 13.14 mg NO3- as N/l), nitrite (0.06 - 6.78 mg NO2- as N/l), orthophosphate (0.07-4.81 mg PO43- as P/l), DO (1.24 - 11.22 mg/l) and turbidity (2.04 -159.06 NTU). Temperature, COD and nitrite varied significantly with season (P < 0.05), while pH, EC, salinity, TDS, COD, and nitrate all varied significantly with sampling site (P < 0.01; P < 0.05). In the rural wastewater treatment facility, free-living Vibrio densities varied from 0 to 3.45 × 101 cfu ml-1, while the plankton-associated Vibrio densities vary with plankton sizes as follows: 180 μm (0 – 4.50 × 103 cfu ml-1); 60 μm (0 – 4.86 × 103 cfu ml-1); 20 μm (0 – 1.9 × 105 cfu ml-1). The seasonal variations in the Vibrio densities in the 180 and 60 μm plankton size samples were significant (P < 0.05), while the 20 μm plankton size and free-living vibrios densities were not. Molecular confirmation of the presumptive vibrios isolates revealed V. fluvialis (36.5 percent), as the predominant species, followed by V. vulnificus (34.6 percent), and V. parahaemolyticus (23.1 percent), and V. metschnikovii (5.8 percent) (detected using only API 20 NE), suggesting high incidence of pathogenic Vibrio species in the final effluent of the wastewater facility. Correlation analysis suggested that the concentration of Vibrio species correlated negatively with salinity and temperature (P < 0.001 and P < 0.002 respectively) as well as with pH and turbidity (P < 0.001), in the final effluent. Population density of total Vibrio ranged from 2.1 × 101 to 4.36 × 104 cfu ml-1 and from 2.80 ×101 to 1.80 × 105 cfu ml-1 for the sub-urban and urban communities treatment facilities respectively. Vibrio species associated with 180 μm, 60 μm, and 20 μm plankton sizes, were observed at densities of 0 - 1.36 × 103 cfu ml-1, 0 - 8.40 × 102 cfu ml-1 and 0 - 6.80 × 102 cfu ml-1 respectively at the sub-urban community‘s WWTP. In the urban community, counts of culturable vibrios ranged from 0 - 2.80 × 102 cfu ml-1 (180 μm); 0 - 6.60 × 102 cfu ml-1 (60 μm) and 0 -1.80 × 103 cfu ml-1 (20 μm). Abundance of free-living Vibrio species varied between 0 and the orders of 102 and 103 cfu ml-1 in the sub-urban and urban communities WWTPs respectively. Molecular confirmation of the presumptive vibrios isolates revealed the presence of V. fluvialis (41.38 percent), V. vulnificus (34.48 percent), and V. parahaemolyticus (24.14 percent) in the sub-urban community effluents. In the urban community V. fluvialis (40 percent), V. vulnificus (36 percent), and V. parahaemolyticus (24 percent) were detected. There was no significant correlation between Vibrio abundance and season, either as free-living or plankton-associated entities, while Vibrio species abundance correlated positively with temperature (r = 0.565; P < 0.01), salinity and dissolved oxygen (P < 0.05). Turbidity and pH showed significant seasonal variation (P < 0.05) in both locations. The Vibrio strains showed the typical multi-antibiotic-resistance of an SXT element. They were resistant to sulfamethoxazole (Sul), trimethoprim (Tmp), cotrimoxazole (Cot), chloramphenicol (Chl) and streptomycin (Str), as well as other antibiotics such as ampicillin (Amp), penicillin (Pen), erythromycin (Ery), tetracycline (Tet), nalidixic acid (Nal), and gentamicin (Gen). The antibiotic resistance genes detected includes dfr18 and dfrA1 for trimethoprim; tetA, strB, floR, sul2 blaP1, for tetracycline, streptomycin, chloramphenicol, sulfamethoxazole and β-lactams respectively. A number of these genes were only recently described from clinical isolates, demonstrating genetic exchange between clinical and environmental Vibrio species. This study revealed that there was an adverse impact on the physicochemical characteristics of the receiving watershed as a result of the discharge of inadequately treated effluents from the wastewater treatment facilities. The occurrence of Vibrio species as plankton-associated entities confirms the role of plankton as potential reservoir for this pathogen. Also the treated final effluents are reservoirs of various antibiotics resistance genes. This could pose significant health and environmental risk to the biotic component of the environment including communities that rely on the receiving water for domestic purposes and may also affect the health status of the aquatic milieu in the receiving water. There is need for consistent monitoring programme by appropriate regulatory agencies to ensure compliance of the wastewater treatment facilities to regulatory effluent quality standards.
- Full Text:
- Date Issued: 2010
- Authors: Igbinosa, Etinosa Ogbomoede
- Date: 2010
- Subjects: Vibrio -- South Africa -- Eastern Cape , Water -- Fluoridation -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11267 , http://hdl.handle.net/10353/245 , Vibrio -- South Africa -- Eastern Cape , Water -- Fluoridation -- South Africa -- Eastern Cape , Water -- Pollution -- South Africa -- Eastern Cape , Effluent quality -- South Africa -- Eastern Cape , Water -- Purification -- South Africa -- Eastern Cape
- Description: Vibrio infections remain a serious threat to public health. In the last decade, Vibrio disease outbreaks have created a painful awareness of the personal, economic, societal, and public health costs associated with the impact of contaminated water in the aquatic milieu. This study was therefore designed to assess the prevalence of Vibrio pathogens in the final effluents of wastewater treatment plants (WWTPs) in the Eastern Cape Province, as well as their abilities to survive the treatment processes of the activated sludge system either as free cells or as plankton-associated entities in relation to the physicochemical qualities of the effluents. Three wastewater treatment facilities were selected to represent typical urban, sub-urban and rural communities, and samples were collected monthly from August 2007 to July 2008 from the final effluent, discharge point, 500 meter upstream and downstream of the discharge points and analysed for physicochemical parameters, Vibrio pathogens prevalence and their antibiogram characteristics using both culture based and molecular techniques. Physicochemical parameters measured include pH, temperature, electrical conductivity, salinity, turbidity, total dissolved solid (TDS), dissolved oxygen (DO), chemical oxygen demand (COD), nitrate, nitrite and orthophosphate levels. Unacceptably high levels of the assayed parameters were observed in many cases for COD (<10 - 1180 mg/l), nitrate (0.08 - 13.14 mg NO3- as N/l), nitrite (0.06 - 6.78 mg NO2- as N/l), orthophosphate (0.07-4.81 mg PO43- as P/l), DO (1.24 - 11.22 mg/l) and turbidity (2.04 -159.06 NTU). Temperature, COD and nitrite varied significantly with season (P < 0.05), while pH, EC, salinity, TDS, COD, and nitrate all varied significantly with sampling site (P < 0.01; P < 0.05). In the rural wastewater treatment facility, free-living Vibrio densities varied from 0 to 3.45 × 101 cfu ml-1, while the plankton-associated Vibrio densities vary with plankton sizes as follows: 180 μm (0 – 4.50 × 103 cfu ml-1); 60 μm (0 – 4.86 × 103 cfu ml-1); 20 μm (0 – 1.9 × 105 cfu ml-1). The seasonal variations in the Vibrio densities in the 180 and 60 μm plankton size samples were significant (P < 0.05), while the 20 μm plankton size and free-living vibrios densities were not. Molecular confirmation of the presumptive vibrios isolates revealed V. fluvialis (36.5 percent), as the predominant species, followed by V. vulnificus (34.6 percent), and V. parahaemolyticus (23.1 percent), and V. metschnikovii (5.8 percent) (detected using only API 20 NE), suggesting high incidence of pathogenic Vibrio species in the final effluent of the wastewater facility. Correlation analysis suggested that the concentration of Vibrio species correlated negatively with salinity and temperature (P < 0.001 and P < 0.002 respectively) as well as with pH and turbidity (P < 0.001), in the final effluent. Population density of total Vibrio ranged from 2.1 × 101 to 4.36 × 104 cfu ml-1 and from 2.80 ×101 to 1.80 × 105 cfu ml-1 for the sub-urban and urban communities treatment facilities respectively. Vibrio species associated with 180 μm, 60 μm, and 20 μm plankton sizes, were observed at densities of 0 - 1.36 × 103 cfu ml-1, 0 - 8.40 × 102 cfu ml-1 and 0 - 6.80 × 102 cfu ml-1 respectively at the sub-urban community‘s WWTP. In the urban community, counts of culturable vibrios ranged from 0 - 2.80 × 102 cfu ml-1 (180 μm); 0 - 6.60 × 102 cfu ml-1 (60 μm) and 0 -1.80 × 103 cfu ml-1 (20 μm). Abundance of free-living Vibrio species varied between 0 and the orders of 102 and 103 cfu ml-1 in the sub-urban and urban communities WWTPs respectively. Molecular confirmation of the presumptive vibrios isolates revealed the presence of V. fluvialis (41.38 percent), V. vulnificus (34.48 percent), and V. parahaemolyticus (24.14 percent) in the sub-urban community effluents. In the urban community V. fluvialis (40 percent), V. vulnificus (36 percent), and V. parahaemolyticus (24 percent) were detected. There was no significant correlation between Vibrio abundance and season, either as free-living or plankton-associated entities, while Vibrio species abundance correlated positively with temperature (r = 0.565; P < 0.01), salinity and dissolved oxygen (P < 0.05). Turbidity and pH showed significant seasonal variation (P < 0.05) in both locations. The Vibrio strains showed the typical multi-antibiotic-resistance of an SXT element. They were resistant to sulfamethoxazole (Sul), trimethoprim (Tmp), cotrimoxazole (Cot), chloramphenicol (Chl) and streptomycin (Str), as well as other antibiotics such as ampicillin (Amp), penicillin (Pen), erythromycin (Ery), tetracycline (Tet), nalidixic acid (Nal), and gentamicin (Gen). The antibiotic resistance genes detected includes dfr18 and dfrA1 for trimethoprim; tetA, strB, floR, sul2 blaP1, for tetracycline, streptomycin, chloramphenicol, sulfamethoxazole and β-lactams respectively. A number of these genes were only recently described from clinical isolates, demonstrating genetic exchange between clinical and environmental Vibrio species. This study revealed that there was an adverse impact on the physicochemical characteristics of the receiving watershed as a result of the discharge of inadequately treated effluents from the wastewater treatment facilities. The occurrence of Vibrio species as plankton-associated entities confirms the role of plankton as potential reservoir for this pathogen. Also the treated final effluents are reservoirs of various antibiotics resistance genes. This could pose significant health and environmental risk to the biotic component of the environment including communities that rely on the receiving water for domestic purposes and may also affect the health status of the aquatic milieu in the receiving water. There is need for consistent monitoring programme by appropriate regulatory agencies to ensure compliance of the wastewater treatment facilities to regulatory effluent quality standards.
- Full Text:
- Date Issued: 2010
Synergistic potententials and isolation of bioactive compounds from the extracts of two helichrysum species indigenous to the Eastern Cape province
- Authors: Aiyegoro, Olayinka Ayobami
- Date: 2010
- Subjects: Helichrysum -- South Africa -- Eastern Cape , Antibiotics , Antioxidants , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11268 , http://hdl.handle.net/10353/250 , Helichrysum -- South Africa -- Eastern Cape , Antibiotics , Antioxidants , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape
- Description: Helichrysum longifolium and H. pedunculatum belong to the Astereceae family and are used extensively in folkloric medicine in South Africa to manage stress-related ailments and as dressings for wounds normally encountered in circumcision rites, bruises, cuts and sores. The in vitro antibacterial time-kill studies, the synergistic potentials, the phytochemical screenings and antioxidant potentials as well as the isolation of the bioactive compounds from the extracts of these two plants were carried out in this study. The in vitro antibacterial activities and time kill regimes of crude extracts of H. pedunculatum was assessed. The extracts was active against both Gram positive and Gram negative bacteria tested at a concentration of 10 mg/ml. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.1 – 35 mg/ml. The average log reduction in viable cell count in time kill assay ranged between 0.17 Log10 to 6.37 Log10 cfu/ml after 6 h of interaction, and between 0.14 Log10 and 6.99 Log10 cfu/ml after 12 h interaction in 1 × MIC and 2 × MIC of the extract. The effect of the aqueous extract was only bacteriostatic on both reference and environmental strains and the clinical isolates were outrightly resistant to aqueous extract. This is worrisome and this could be one reason why, there is an incidence of high death rate resulting from circumcision wounds infection even after treating such wounds with H. pedunculatum leaf. In vitro antibacterial time kill studies of extracts of H. longifolium was assessed. All test bacteria were susceptible to the methanol extract, while none was susceptible to the aqueous extract. Two of the test bacteria were susceptible to the ethyl acetate extract, while ten and seven were susceptible to the acetone and chloroform extracts respectively at the test concentration of 5 mg/ml. The minimum inhibitory concentrations (MICs) ranged between 0.1 and 5.0 mg/ml, while minimum bactericidal concentrations (MBCs) ranged between 1.0 and >5 mg/ml for all the extracts. Average log reductions in viable cell counts for all the extracts ranged between 0.1 Log10 and 7.5 Log10 cfu/ml after 12 h interaction at 1 × MIC and 2 × MIC. Most of the extracts were rapidly bactericidal at 2 × MIC achieving a complete elimination of most of the test organisms within 12 h exposure time. The effect of combinations of the crude extracts of H. pedunculatum leaves and eight antibiotics was investigated by means of checkerboard and time-kill methods. In the checkerboard method, synergies of between 45.83-56.81 percent were observed and this is independent of Gram reaction, with combinations in the aqueous extract yielding largely antagonistic interactions (18.75 percent). The time kill assay also detected synergy that is independent of Gram reaction with a ≥ 3Log10 potentiation of the bactericidal activity of the test antibiotics. We conclude that the crude leaf extracts of H. pedunculatum could be potential source of broad spectrum antibiotics resistance modulating compounds. The interactions between crude extracts of H. longifolium in combination with six first-line antibiotics using both the time-kill and the checkerboard methods were carried out. The time-kill method revealed the highest bactericidal activity exemplified by a 6.7 Log10 reduction in cell density against Salmonella sp. when the extract and Penicillin G are combined at ½ × MIC. Synergistic response constituted about 65 percent, while indifference and antagonism constituted about 28.33 percent and 6.67 percent in the time kill assay, respectively. The checkerboard method also revealed that the extracts improved bactericidal effects of the antibiotics. About 61.67 percent of all the interactions were synergistic, while indifference interactions constituted about 26.67 percent and antagonistic interactions was observed in approximately 11.66 percent. The in vitro antioxidant property and phytochemical constituents of the aqueous crude leaf extracts of H. longifolium and H. pedunculatum was investigated. The scavenging activity on superoxide anions, DPPH, H2O2, NO and ABTS; and the reducing power were determined, as well as the flavonoid, proanthocyanidin and phenolic contents of the extracts. The extracts exhibited scavenging activity in all radicals tested due to the presence of relatively high total phenol and flavonoids contents in the extracts. Our findings suggest that H. longifolium and H. pedunculatum are endowed with antioxidant phytochemicals and could serve as a base for future drugs. Bioactivity-guided fractionation of the leaves of H. longifolium and H. pedunculatum yielded two known compounds. From the n-hexane fraction of H. longifolium a compound was isolated (Stigmasterol) and from the ethyl acetate fraction of H. pedunculatum another compound (β-sitosterol) was isolated. The compounds were isolated and identified using various techniques. The antimicrobial, anti-inflammatory, antioxidant, analgesic and anti-pyretic activities of these compounds have been reported in literatures. In general, the experiments and tests conducted in this study appear to have justified the folkloric medicinal uses of H. longifolium and H. pedunculatum for the treatment of stress related ailments and wound infections and make a substantial contribution to the knowledge base of the use of herbal medicine for the treatment of the microbial infections.
- Full Text:
- Date Issued: 2010
- Authors: Aiyegoro, Olayinka Ayobami
- Date: 2010
- Subjects: Helichrysum -- South Africa -- Eastern Cape , Antibiotics , Antioxidants , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11268 , http://hdl.handle.net/10353/250 , Helichrysum -- South Africa -- Eastern Cape , Antibiotics , Antioxidants , Medicinal plants -- South Africa -- Eastern Cape , Traditional medicine -- South Africa -- Eastern Cape
- Description: Helichrysum longifolium and H. pedunculatum belong to the Astereceae family and are used extensively in folkloric medicine in South Africa to manage stress-related ailments and as dressings for wounds normally encountered in circumcision rites, bruises, cuts and sores. The in vitro antibacterial time-kill studies, the synergistic potentials, the phytochemical screenings and antioxidant potentials as well as the isolation of the bioactive compounds from the extracts of these two plants were carried out in this study. The in vitro antibacterial activities and time kill regimes of crude extracts of H. pedunculatum was assessed. The extracts was active against both Gram positive and Gram negative bacteria tested at a concentration of 10 mg/ml. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.1 – 35 mg/ml. The average log reduction in viable cell count in time kill assay ranged between 0.17 Log10 to 6.37 Log10 cfu/ml after 6 h of interaction, and between 0.14 Log10 and 6.99 Log10 cfu/ml after 12 h interaction in 1 × MIC and 2 × MIC of the extract. The effect of the aqueous extract was only bacteriostatic on both reference and environmental strains and the clinical isolates were outrightly resistant to aqueous extract. This is worrisome and this could be one reason why, there is an incidence of high death rate resulting from circumcision wounds infection even after treating such wounds with H. pedunculatum leaf. In vitro antibacterial time kill studies of extracts of H. longifolium was assessed. All test bacteria were susceptible to the methanol extract, while none was susceptible to the aqueous extract. Two of the test bacteria were susceptible to the ethyl acetate extract, while ten and seven were susceptible to the acetone and chloroform extracts respectively at the test concentration of 5 mg/ml. The minimum inhibitory concentrations (MICs) ranged between 0.1 and 5.0 mg/ml, while minimum bactericidal concentrations (MBCs) ranged between 1.0 and >5 mg/ml for all the extracts. Average log reductions in viable cell counts for all the extracts ranged between 0.1 Log10 and 7.5 Log10 cfu/ml after 12 h interaction at 1 × MIC and 2 × MIC. Most of the extracts were rapidly bactericidal at 2 × MIC achieving a complete elimination of most of the test organisms within 12 h exposure time. The effect of combinations of the crude extracts of H. pedunculatum leaves and eight antibiotics was investigated by means of checkerboard and time-kill methods. In the checkerboard method, synergies of between 45.83-56.81 percent were observed and this is independent of Gram reaction, with combinations in the aqueous extract yielding largely antagonistic interactions (18.75 percent). The time kill assay also detected synergy that is independent of Gram reaction with a ≥ 3Log10 potentiation of the bactericidal activity of the test antibiotics. We conclude that the crude leaf extracts of H. pedunculatum could be potential source of broad spectrum antibiotics resistance modulating compounds. The interactions between crude extracts of H. longifolium in combination with six first-line antibiotics using both the time-kill and the checkerboard methods were carried out. The time-kill method revealed the highest bactericidal activity exemplified by a 6.7 Log10 reduction in cell density against Salmonella sp. when the extract and Penicillin G are combined at ½ × MIC. Synergistic response constituted about 65 percent, while indifference and antagonism constituted about 28.33 percent and 6.67 percent in the time kill assay, respectively. The checkerboard method also revealed that the extracts improved bactericidal effects of the antibiotics. About 61.67 percent of all the interactions were synergistic, while indifference interactions constituted about 26.67 percent and antagonistic interactions was observed in approximately 11.66 percent. The in vitro antioxidant property and phytochemical constituents of the aqueous crude leaf extracts of H. longifolium and H. pedunculatum was investigated. The scavenging activity on superoxide anions, DPPH, H2O2, NO and ABTS; and the reducing power were determined, as well as the flavonoid, proanthocyanidin and phenolic contents of the extracts. The extracts exhibited scavenging activity in all radicals tested due to the presence of relatively high total phenol and flavonoids contents in the extracts. Our findings suggest that H. longifolium and H. pedunculatum are endowed with antioxidant phytochemicals and could serve as a base for future drugs. Bioactivity-guided fractionation of the leaves of H. longifolium and H. pedunculatum yielded two known compounds. From the n-hexane fraction of H. longifolium a compound was isolated (Stigmasterol) and from the ethyl acetate fraction of H. pedunculatum another compound (β-sitosterol) was isolated. The compounds were isolated and identified using various techniques. The antimicrobial, anti-inflammatory, antioxidant, analgesic and anti-pyretic activities of these compounds have been reported in literatures. In general, the experiments and tests conducted in this study appear to have justified the folkloric medicinal uses of H. longifolium and H. pedunculatum for the treatment of stress related ailments and wound infections and make a substantial contribution to the knowledge base of the use of herbal medicine for the treatment of the microbial infections.
- Full Text:
- Date Issued: 2010
Prevalence of Escherichia coli O157:H7 in water and meat and meat products and vegetables sold in the Eastern Cape Province of South Africa and its impact on the diarrhoeic conditions of HIV/AIDS patients
- Authors: Abong'o, Benard Omondi
- Date: 2008
- Subjects: Foodborne diseases , Diarrhea , Escherichia coli , HIV infections , AIDS (Disease) , Bacterial diseases
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11263 , http://hdl.handle.net/10353/87 , Foodborne diseases , Diarrhea , Escherichia coli , HIV infections , AIDS (Disease) , Bacterial diseases
- Description: Water and food borne Escherichia coli O157:H7 could be one of the pathogens posing high health risk to patients suffering from Acquired Immunodeficiency Syndrome (AIDS) because of its incrimination in diarrhoea cases in AIDS patients. The present study, which was conducted between March 2005 and August 2006, investigated the prevalence of E. coli O157:H7 in water, meat and meat products and vegetables and its impact on diarrhoeic conditions of confirmed and non-confirmed HIV/AIDS patients in the Amathole District in the Eastern Cape Province of South Africa. The water samples used in the study were obtained from stand pipes supplying treated drinking water to communities residing in Fort Beaufort, Alice, Dimbaza and Mdantsane whereas borehole waters were sampled from Ngwenya and Kwasaki. The meat and meat products and vegetable samples were purchased from shops, butcheries, supermarkets and open air markets in Fort Beaufort, Alice and Mdantsane. The stool swabs used in the study were obtained from HIV/AIDS and outpatient clinics at Frere Hospital in East London. A total of 180 each of water, meat and meat products and vegetable samples and another 360 stool samples were analyzed for E. coli O157:H7. Presumptive E. coli O157 was isolated from the samples by culture-based methods and confirmed using Polymerase Chain Reaction techniques. Anti-biogram as well as risk assessment were also carried out using standard methods. The viable counts of presumptive E. coli O157 for water samples ranged between 3.3 × 104 and 1.71 × 105 CFU/ml, and between 1.8 × 104 and 5.04 × 106 CFU/g for meat and meat products, whereas those for vegetables ranged between 1.3 × 103 and 1.6 × 106 CFU/g. The counts of presumptive E. coli O157 for the water and vegetable samples were not significantly different whereas those for meat and meat products were found to be significantly different (P ≤ 0.05). The prevalence rates of presumptive E coli O157 in meat and meat products was 35.55 percent (64/180), and 25.55 percent (46/180) and 21.66 percent (39/180) for water and vegetables respectively. Prevalence of presumptive E. coli O157 in the stool samples of HIV/AIDS patients was 36.39 percent (131/360), of which 56.5 percent (74/131) and 43.5 percent (57/131) were from stools of confirmed and non-confirmed HIV/AIDS patients, respectively. Molecular analysis of representative presumptive E. coli O157 indicated that 10.29 percent (4/39) of vegetables; 14.81 percent (4/27) of water and 38.46 percent (5/13) of meat and meat products carried E. coli O157:H7. Also 36 percent (9/25) and 17.24 percent (5/29) of the stool samples were positive for E. coli O157:H7. Antimicrobial susceptibility profile revealed that all of the E. coli O157:H7 isolated from water, meat and meat products and vegetables as well as those isolated from stools of confirmed and non-confirmed HIV/AIDS patients were resistant (R) to gentamycin and erythromycin. However, 75 percent (20/27) of these isolates were resistant (R) to ampicillin and tetracycline whereas approximately 25 percent (6/27) were resistant (R) to nalidixic acid, ceftriaxone, and chloramphenicol. All the isolates (27/27) were susceptible (S) to amikacin. Probability of risk of E. coli O157:H7 infection was high for confirmed HIV/AIDS patients than for the non-confirmed HIV/AIDS patients. Estimated probability of risk of E. coli O157:H7 due to ingestion of water was 1.00 for 100 confirmed and non-confirmed HIV/AIDS patients. Risk due to meat and meat products was estimated at 0.27 and 0.20 and for vegetables at 0.21 and 0.15 per 100 confirmed and non-confirmed HIV/AIDS patients. The findings of this study predicted a possible link between E. coli O157:H7 isolated from drinking water, meat and meat products and vegetables and diarrhoeic conditions in both confirmed and non-confirmed HIV/AIDS patients, and concludes that confirmed HIV/AIDS patients can be at higher risk of contracting water and food borne E. coli O157:H7 than nonconfirmed HIV/AIDS patients. It is thus recommended that proper water treatment and food handling, maximum food and water safety and sanitation as well as personal body hygiene should be maintained, in order to prevent E. coli O157:H7 infections. Education initiatives and active surveillance of E. coli O157:H7 should be taken by all the stake-holders working directly or indirectly towards ensuring enduring sound public health.
- Full Text:
- Date Issued: 2008
- Authors: Abong'o, Benard Omondi
- Date: 2008
- Subjects: Foodborne diseases , Diarrhea , Escherichia coli , HIV infections , AIDS (Disease) , Bacterial diseases
- Language: English
- Type: Thesis , Doctoral , PhD (Microbiology)
- Identifier: vital:11263 , http://hdl.handle.net/10353/87 , Foodborne diseases , Diarrhea , Escherichia coli , HIV infections , AIDS (Disease) , Bacterial diseases
- Description: Water and food borne Escherichia coli O157:H7 could be one of the pathogens posing high health risk to patients suffering from Acquired Immunodeficiency Syndrome (AIDS) because of its incrimination in diarrhoea cases in AIDS patients. The present study, which was conducted between March 2005 and August 2006, investigated the prevalence of E. coli O157:H7 in water, meat and meat products and vegetables and its impact on diarrhoeic conditions of confirmed and non-confirmed HIV/AIDS patients in the Amathole District in the Eastern Cape Province of South Africa. The water samples used in the study were obtained from stand pipes supplying treated drinking water to communities residing in Fort Beaufort, Alice, Dimbaza and Mdantsane whereas borehole waters were sampled from Ngwenya and Kwasaki. The meat and meat products and vegetable samples were purchased from shops, butcheries, supermarkets and open air markets in Fort Beaufort, Alice and Mdantsane. The stool swabs used in the study were obtained from HIV/AIDS and outpatient clinics at Frere Hospital in East London. A total of 180 each of water, meat and meat products and vegetable samples and another 360 stool samples were analyzed for E. coli O157:H7. Presumptive E. coli O157 was isolated from the samples by culture-based methods and confirmed using Polymerase Chain Reaction techniques. Anti-biogram as well as risk assessment were also carried out using standard methods. The viable counts of presumptive E. coli O157 for water samples ranged between 3.3 × 104 and 1.71 × 105 CFU/ml, and between 1.8 × 104 and 5.04 × 106 CFU/g for meat and meat products, whereas those for vegetables ranged between 1.3 × 103 and 1.6 × 106 CFU/g. The counts of presumptive E. coli O157 for the water and vegetable samples were not significantly different whereas those for meat and meat products were found to be significantly different (P ≤ 0.05). The prevalence rates of presumptive E coli O157 in meat and meat products was 35.55 percent (64/180), and 25.55 percent (46/180) and 21.66 percent (39/180) for water and vegetables respectively. Prevalence of presumptive E. coli O157 in the stool samples of HIV/AIDS patients was 36.39 percent (131/360), of which 56.5 percent (74/131) and 43.5 percent (57/131) were from stools of confirmed and non-confirmed HIV/AIDS patients, respectively. Molecular analysis of representative presumptive E. coli O157 indicated that 10.29 percent (4/39) of vegetables; 14.81 percent (4/27) of water and 38.46 percent (5/13) of meat and meat products carried E. coli O157:H7. Also 36 percent (9/25) and 17.24 percent (5/29) of the stool samples were positive for E. coli O157:H7. Antimicrobial susceptibility profile revealed that all of the E. coli O157:H7 isolated from water, meat and meat products and vegetables as well as those isolated from stools of confirmed and non-confirmed HIV/AIDS patients were resistant (R) to gentamycin and erythromycin. However, 75 percent (20/27) of these isolates were resistant (R) to ampicillin and tetracycline whereas approximately 25 percent (6/27) were resistant (R) to nalidixic acid, ceftriaxone, and chloramphenicol. All the isolates (27/27) were susceptible (S) to amikacin. Probability of risk of E. coli O157:H7 infection was high for confirmed HIV/AIDS patients than for the non-confirmed HIV/AIDS patients. Estimated probability of risk of E. coli O157:H7 due to ingestion of water was 1.00 for 100 confirmed and non-confirmed HIV/AIDS patients. Risk due to meat and meat products was estimated at 0.27 and 0.20 and for vegetables at 0.21 and 0.15 per 100 confirmed and non-confirmed HIV/AIDS patients. The findings of this study predicted a possible link between E. coli O157:H7 isolated from drinking water, meat and meat products and vegetables and diarrhoeic conditions in both confirmed and non-confirmed HIV/AIDS patients, and concludes that confirmed HIV/AIDS patients can be at higher risk of contracting water and food borne E. coli O157:H7 than nonconfirmed HIV/AIDS patients. It is thus recommended that proper water treatment and food handling, maximum food and water safety and sanitation as well as personal body hygiene should be maintained, in order to prevent E. coli O157:H7 infections. Education initiatives and active surveillance of E. coli O157:H7 should be taken by all the stake-holders working directly or indirectly towards ensuring enduring sound public health.
- Full Text:
- Date Issued: 2008
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