Theses and Dissertations

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A comparative study of the various pretreatment techniques for bioethanol production from Kikuyu Grass
(University of Fort Hare, 2013-12) Mbande, Zukiswa Coceka
Bio-ethanol is by far the most widely used bio-fuel for transportation worldwide. Production of bio-ethanol from biomass is one way to reduce both consumption of crude oil and environmental pollution. There are mainly two processes involved in the conversion of lignocellulosic materials to ethanol production: hydrolysis of cellulose in the lignocellulosic biomass to produce reducing sugars and fermentation of the sugars to ethanol. However, pretreatment is an essential step before the hydrolysis of biomass and subsequent production of bio-ethanol. The current study was focused on four different chemical methods for the pretreatment of Kikuyu grass using mild temperatures. The temperature used was 100°C for various periods compared.
Antibacterial and phytochemical studies of selected South African honeys on clinical isolates of Helicobacter pylori
(University of Fort Hare, 2012) Manyi-Loh, Christy E; Clarke, A; Ndip, R
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
Antibacterial properties of the methanol extract of helichrysum pedunculatum
(University of Fort Hare, 2008) Ncube, Nqobile S; Okoh, Anthony I; Afolayan, A J
The methanol extract of Helichrisum pedunculatum was screened for antimicrobial activity up to a concentration of 5 mg/ml using the agar dilution technique. A number of test bacterial isolates, comprising both Gram negative and Gram positive organisms were susceptible to the crude extract of the plant. The minimum inhibitory concentrations (MICs) of the extract ranged between 1 and 5 mg/ml for the susceptible organisms. The MICs of the selected antibiotics, chloramphenicol and penicillin, ranged between 2 and 4 mg/L, and 2 and 32 mg/L respectively against Bacillus cereus, Proteus vulgaris and Staphylococcus aureus OKOH1. Bactericidal activity was determined by the time kill assay. The methanol extract of the plant was not bactericidal at 1 × MIC for B. cereus, P. vulgaris and Staph. aureus OKOH1. At 2 × MIC the extract was bacteriostatic against B. cereus but bactericidal against P. vulgaris and Staph. aureus OKOH1. Combination studies were done at 1/2 × MIC, 1 × MIC and 2 × MIC of the plant extract with 1 × MIC of the antibiotics. Combinations of the plant extract and chloramphenicol resulted in mostly indifferent interactions against P. vulgaris and Staph. aureus OKOH1 but synergistic interactions at higher concentration of the plant extract for B. cereus. Penicillin combinations gave synergistic interactions at lower concentrations of the plant for P.vulgaris and Staph. aureus OKOH1 but was mostly indifferent for B. cereus.
Antibiogram profiling of Escherichia coli pathotypes isolated from Kat River and Fort Beaufort abstraction water
(University of Fort Hare, 2014) Nontongana, Nolonwabo; Okoh, Anthony I
Escherichia coli (E. coli) is a widespread species that includes a broad variety of strains, ranging from highly pathogenic strains causing worldwide outbreaks of severe disease to virulent isolates which are part of the normal intestinal flora or which are well characterized and safe laboratory strains. The pathogenicity of a given E. coli strain is mainly determined by specific virulence factors which include adhesins, invasins, toxins and capsule. The aim of this study was to evaluate the prevalence and antibiogram profiles of E. coli pathotypes previously isolated from Kat River and Fort Beaufort abstraction water. A total of 171 E. coli isolates showed at least one pathogenic determinant among the isolated 278 E. coli. The other 107 isolates were negative for the tested virulence genes. All 278 presumptive isolates tested positive for the UidA gene, and were therefore classified as non-categorized pathogenic E. coli. The 171 pathogenic isolates had at least one characteristic gene of pathogenic E. coli and were identified and classified as enteropathogenic E. coli (6%), enterotoxigenicE. coli (131), uropathogenic E. coli (6), neonatal meningitis E. coli (14), diffusely adherent E. coli (1) and enterohaemrrhagic E. coli (1). Interestingly, no virulence genes were detected for the enteroinvasive E. coli and the enteroaggregative E. coli. The antibiotic resistance profiles for all isolates that were identified as E. coli showed 100% resistance to penicillin G, 98% resistance to ampicillin, 38% resistance to trimethoprim-sulphamethoxazole and 8% resistance to streptomycin. Multiple antibacterial resistance (MAR) was also observed, where 44% of the isolates were resistant to three antibiotics and 8% resistant to four antibiotics. The results of this study showed the Kat River and Fort Beaufort abstraction water are reservoirs of pathogenic strains of E. coli which harbour antibiotic resistance determinants that can cause serious health risks to the people in the surrounding communities.
Antibiogram profiling of Escherichia Coli pathotypes isolated from Kat River and Fort Beaufort abstraction water.
(University of Fort Hare, 2014) Nontongana, Nolonwabo
Escherichia coli (E. coli) is a widespread species that includes a broad variety of strains, ranging from highly pathogenic strains causing worldwide outbreaks of severe disease to virulent isolates which are part of the normal intestinal flora or which are well characterized and safe laboratory strains. The pathogenicity of a given E. coli strain is mainly determined by specific virulence factors which include adhesins, invasins, toxins and capsule. The aim of this study was to evaluate the prevalence and antibiogram profiles of E. coli pathotypes previously isolated from Kat River and Fort Beaufort abstraction water. A total of 171 E. coli isolates showed at least one pathogenic determinant among the isolated 278 E. coli. The other 107 isolates were negative for the tested virulence genes. All 278 presumptive isolates tested positive for the UidA gene, and were therefore classified as non-categorized pathogenic E. coli. The 171 pathogenic isolates had at least one characteristic gene of pathogenic E. coli and were identified and classified as enteropathogenic E. coli (6%), enterotoxigenicE. coli (131), uropathogenic E. coli (6), neonatal meningitis E. coli (14), diffusely adherent E. coli (1) and enterohaemrrhagic E. coli (1). Interestingly, no virulence genes were detected for the enteroinvasive E. coli and the enteroaggregative E. coli .The antibiotic resistance profiles for all isolates that were identified as E. coli showed 100% resistance to penicillin G, 98% resistance to ampicillin, 38% resistance to trimethoprim-sulphamethoxazole and 8% resistance to streptomycin. Multiple antibacterial resistance (MAR) was also observed, where 44% of the isolates were resistant to three antibiotics and 8% resistant to four antibiotics. The results of this study showed the Kat River and Fort Beaufort abstraction water are reservoirs of pathogenic strains of E. coli which harbour antibiotic resistance determinants that can cause serious health risks to the people in the surrounding communities.
Assessment of antibacterial potentials of Garcinia Kola seed extracts and their interactions with antibiotics
(University of Fort Hare, 2007) Sibanda, Thulani; Okoh, Anthony I
The antibacterial potency of the extracts of the seed of Garcinia kola (bitter kola) was investigated in this study against a panel of referenced, environmental and clinical bacterial strains. The killing rates of the active extract as well as their potential for combination antibacterial therapy with standard antibiotics were also elucidated using standard procedures. The aqueous and acetone extracts of the seed were screened for activity against 27 bacterial isolates. The aqueous extract exhibited activity mainly against Gram positive organisms with Minimum inhibitory concentration (MIC) values ranging from 5 mgml-1 – 20 mgml-1, while the acetone extract showed activity against both Gram negative and Gram positive organisms with MIC values ranging from 10 mgml-1 - 0.156 mgml-1. The acetone extract also showed rapid bactericidal activity against Staphylococcus aureus ATCC 6538 with a 3.097 Log10 reduction in counts within 4 hours at 0.3125 mgml-1 and a 1.582 Log10 reduction against Proteus vulgaris CSIR 0030 at 5 mgml-1 after 1 hour. In addition, the aqueous, methanol and acetone extracts of the seeds also exhibited activity against four clinical strains of Staphylococcus isolated from wound sepsis specimens. The MIC values for the aqueous extract were 10 mgml-1 for all the isolates while the acetone and methanol extracts had lower values ranging from 0.3125 - 0.625 mgml-1. The acetone extract was strongly bactericidal against Staphylococcus aureus OKOH3 resulting in a 2.70 Log10 reduction in counts at 1.25 mgml-1 within 4 hours of exposure and a complete elimination of the organism after 8 hours. The bactericidal vi activity of the same extract against Staphylococcus aureus OKOH1 was weak, achieving only a 2.92 Log10 reduction in counts at 1.25 mgml-1 (4× MIC) in 24 hours. In the test for interactions between the acetone extract of the seeds and antibiotics, synergistic interactions were observed largely against Gram positive organisms using the FIC indices, (indices of 0.52 - 0.875) with combinations against Gram negatives yielding largely antagonistic interactions (indices of 2.0 to 5.0). Synergy (≥ 1000 times or ≥ 3 Log10 potentiation of the bactericidal activity) against both Gram negative and Gram positive organisms was detected by time kill assays mainly involving the antibiotics tetracycline, chloramphenicol, amoxycillin and penicillin G. Combinations involving erythromycin and ciprofloxacin consistently gave antagonistic or indifferent interactions. We conclude that the acetone extract of Garcinia kola seeds possess strong bactericidal activities against both Gram positive and Gram negative organisms and can be therapeutically useful in the treatment of bacterial infections including the problematic staphylococcal wound infections. In addition, the acetone extract can be a potential source of broad spectrum resistance modifying compounds that can potentially improve the performance of antibiotics in the treatment of drug resistant infections.
Assessment of antibiotic production by some marine Streptomyces isolated from the Nahoon Beach
(University of Fort Hare, 2010) Ogunmwonyi, Isoken Nekpen Henrietta
Rapidly emerging strains of bacteria resistant to most advanced antibiotics have become issues of very important public health concern. Research currently directed towards marine actinomycetes presents a vast potential for new compounds that could be able to safely and effectively target resistant species. In this regard, ten putative Streptomyces strains isolated from the Nahoon beach were selected and assessed for antibiotic production and activity against a wide range of bacteria including reference strains, environmental strain and clinical isolates. The ethyl acetate extracts of the putative Streptomyces isolates showed activities against at least 6 and up to 26 of the 32 test bacteria. Inhibition zones were found to range between 9-32 mm diameters at a concentration of 10 mg/ml. The minimum inhibitory concentrations (MICs) of the crude extracts ranged from 0.039 - 10 mg/ml and the least minimum bactericidal concentration (MBC) demonstrated was 0.625 mg/ml against a reference strain Staphylococcus aureus ATCC 6538. Time kill kinetics of all extracts revealed bacteristatic and bactericidal activities. Average Log reductions in viable cell counts for all the extracts ranged from 0.86 Log10 and 3.99 Log10 cfu/ml after 3 h interaction and 0.01 Log10 and 4.86 Log10 after 6 h interaction at MIC, 2 × MIC, 3 × MIC and 4 × MIC concentrations. Most of the extracts were speedily bactericidal at 3 × MIC and 4 × MIC resulting in over 50 % elimination of most of the test bacteria within 3 h and 6 h interaction. The partial characterization of the crude extracts by IR spectral analysis revealed possibility of terpenoid, long chain fatty acids and secondary amine derivatives compounds in the extracts. It is therefore recommended that further investigation should address the relationship between the structure of the active component of the extracts and the broad spectrum activity, as well as a rapid method for large scale production and purification and whether this group of antibiotics has any application in managing human infectious disease..
Assessment of antibiotic production by some marine Streptomyces isolated from the Nahoon Beach
(University of Fort Hare, 2010) Ogunmwonyi, Isoken Nekpen Henrietta; Okoh, I
Rapidly emerging strains of bacteria resistant to most advanced antibiotics have become issues of very important public health concern. Research currently directed towards marine actinomycetes presents a vast potential for new compounds that could be able to safely and effectively target resistant species. In this regard, ten putative Streptomyces strains isolated from the Nahoon beach were selected and assessed for antibiotic production and activity against a wide range of bacteria including reference strains, environmental strain and clinical isolates. The ethyl acetate extracts of the putative Streptomyces isolates showed activities against at least 6 and up to 26 of the 32 test bacteria. Inhibition zones were found to range between 9-32 mm diameters at a concentration of 10 mg/ml. The minimum inhibitory concentrations (MICs) of the crude extracts ranged from 0.039 - 10 mg/ml and the least minimum bactericidal concentration (MBC) demonstrated was 0.625 mg/ml against a reference strain Staphylococcus aureus ATCC 6538. Time kill kinetics of all extracts revealed bacteristatic and bactericidal activities. Average Log reductions in viable cell counts for all the extracts ranged from 0.86 Log10 and 3.99 Log10 cfu/ml after 3 h interaction and 0.01 Log10 and 4.86 Log10 after 6 h interaction at MIC, 2 × MIC, 3 × MIC and 4 × MIC concentrations. Most of the extracts were speedily bactericidal at 3 × MIC and 4 × MIC resulting in over 50 % elimination of most of the test bacteria within 3 h and 6 h interaction. The partial characterization of the crude extracts by IR spectral analysis revealed possibility of terpenoid, long chain fatty acids and secondary amine derivatives compounds in the extracts. It is therefore recommended that further investigation should address the relationship between the structure of the active component of the extracts and the broad spectrum activity, as well as a rapid method for large scale production and purification and whether this group of antibiotics has any application in managing human infectious disease.
Assessment of bioflocculant production by some marine bacteria isolated from the bottom sediment of Algoa Bay
(University of Fort Hare, 2010) Cosa, Sekelwa; Okoh, Anthony I
Several problems concerning the use of conventional synthetic flocculants has necessitated the need for alternative cost effective, safe and efficient bioflocculants from microorganisms inhabiting many environments, particularly those from unusual environments. Hence, this study assessed bioflocculant production by three marine bacteria isolated from the bottom sediment of Algoa Bay in the Eastern Cape Province of South Africa. Analysis of the 16S rDNA sequences led to their identification as Halobacillus sp. Mvuyo, Virgibacillus sp. Rob and Oceanobacillus sp. Pinky. Several factors affecting the production and activity of the bioflocculant(s) were studied. Halobacillus sp. Mvuyo produced bioflocculant optimally with glucose (76%) and ammonium chloride (93%) as sole carbon and nitrogen sources, respectively and at neutral pH and in the presence of Ca2+. On the other hand, Virgibacillus sp. Rob preferred glucose (70.4 %) and iron sulphate (74%) as carbon and nitrogen source respectively; an alkaline pH (12.0) and Fe2+. Oceanobacillus sp. Pinky produced bioflocculant optimally when sucrose (80%) and peptone (72.4 %) were used as carbon and nitrogen source respectively, at neutral pH, and in the presence of Ca2+ cation. The chemical analyses of the partially purified bioflocculants revealed that the bioflocculants produced by Halobacillus sp. Mvuyo and Oceanobacillus sp. Pinky were glycoproteins, while that produced by Virgibacillus sp. Rob was a polysaccharide. We thus conclude that Halobacillus sp. Mvuyo, Virgibacillus sp. Rob and Oceanobacillus sp. Pinky hold promise as producers of new and efficient bioflocculant(s). We recommended development of process conditions for large scale production of the bioflocculants followed by their detailed characterization, as well as pilot scale assessment of the applicability of the purified bioflocculant in water/wastewater treatment and other industrial uses
Assessment of bioflocculant production by two marine bacteria isolated from the bottom sediment of marine Algoa Bay
(University of Fort Hare, 2015) Ntozonke, Ncedo; Prof. Okoh, A I
Bioflocculants are polymers, mostly, of microbial origin which floc out suspended particles from liquid medium. The ability of these biopolymers to remove suspended particles from solutions is termed bioflocculation, and the efficiency of flocculation activities depends on the characteristics of the flocculants. In comparison with conventionally used flocculants, bioflocculants have the advantage of being safe (no toxic effects known), biodegradable and harmlessness to the environment. The study assessed production of bioflocculant by two marine bacteria from the bottom sediment of marine environment. The 16S rDNA was used for identification, and the two bacteria species were identified as Enterococcus hirae and Bacillus thuringiensis. Factors affecting the production and activity of the bioflocculants produced by these two organisms were studied. The bacteria optimally produced bioflocculant with fructose (91.7%) and urea (91%) as sole carbon and nitrogen sources respectively. Mg2+ (87%) and Ca2+ (86%), likewise, served as best cation sources on the production of the bioflocculant at pH 5(93%). Additionally, the flocculating activity of the bioflocculant increased with the addition of Mg2+ (81%) and Na+ (81%), and the highest flocculating activity was at pH 5 of the kaolin clay. The Fourier transform infrared spectroscopy (FTIR) shows that the bioflocculant is a glycoprotein. The second bacterium (Bacillus thuringiensis) produced bioflocculant optimally when the media had mixed nitrogen sources (Urea, ammonium chloride and tryptone (67%)) and glucose (85.65%) as a sole carbon source, also Ca2+ (74.6%) was the best cation that induced the production of bioflocculant. After purification, the bioflocculant flocculated optimally in alkaline pH 12 (81%) in the presence of Mn2+ (73%) and Ca2+ (72.8%). Chemical analysis of the bioflocculant revealed it to be a polysaccharide. Both bioflocculants flocculate efficiently and can be used to replace synthetic flocculants in water treatment, wastewater, in downstream processing, and processing of food and chemicals and other industrial uses of flocculants. Challenges though (i) are to develop conditions for large scale production of the bioflocculant, (ii) to do further characterization of the both bioflocculants (iii) to assess the bioflocculants for treatments of water/wastewater, and to apply it in various industrial processes.
Assessment of bioflocculant production by two marine bacteria isolated from the bottom sediment of marine Algoa Bay
(University of Fort Hare, 2015) Ntozonke, Ncedo; Okoh, Anthony I; Nwodo, Uchechukwu U
Bioflocculants are polymers, mostly, of microbial origin which floc out suspended particles from liquid medium. The ability of these biopolymers to remove suspended particles from solutions is termed bioflocculation, and the efficiency of flocculation activities depends on the characteristics of the flocculants. In comparison with conventionally used flocculants, bioflocculants have the advantage of being safe (no toxic effects known), biodegradable and harmlessness to the environment. The study assessed production of bioflocculant by two marine bacteria from the bottom sediment of marine environment. The 16S rDNA was used for identification, and the two bacteria species were identified as Enterococcus hirae and Bacillus thuringiensis. Factors affecting the production and activity of the bioflocculants produced by these two organisms were studied. The bacteria optimally produced bioflocculant with fructose (91.7%) and urea (91%) as sole carbon and nitrogen sources respectively. Mg2+ (87%) and Ca2+ (86%), likewise, served as best cation sources on the production of the bioflocculant at pH 5(93%). Additionally, the flocculating activity of the bioflocculant increased with the addition of Mg2+ (81%) and Na+ (81%), and the highest flocculating activity was at pH 5 of the kaolin clay. The Fourier transform infrared spectroscopy (FTIR) shows that the bioflocculant is a glycoprotein. The second bacterium (Bacillus thuringiensis) produced bioflocculant optimally when the media had mixed nitrogen sources (Urea, ammonium chloride and tryptone (67%)) and glucose (85.65%) as a sole carbon source, also Ca2+ (74.6%) was the best cation that induced the production of bioflocculant. After purification, the bioflocculant flocculated optimally in alkaline pH 12 (81%) in the presence of Mn2+ (73%) and Ca2+ (72.8%). Chemical analysis of the bioflocculant revealed it to be a polysaccharide. Both bioflocculants flocculate efficiently and can be used to replace synthetic flocculants in water treatment, wastewater, in downstream processing, and processing of food and chemicals and other industrial uses of flocculants. Challenges though (i) are to develop conditions for large scale production of the bioflocculant, (ii) to do further characterization of the both bioflocculants (iii) to assess the bioflocculants for treatments of water/wastewater, and to apply it in various industrial processes.
Assessment of the anti-Listerial properties of Garcinia kola (Heckel) seeds
(University of Fort Hare, 2014) Penduka, Dambudzo; Okoh, Anthony I; Mayekiso, B; Buwa, L
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.
Assessment of the antibacterial properties of n-Hexane extract of Cocos Nucifera and its interactions with some conventional antibiotics
(University of Fort Hare, 2011) Akinyele, Taiwo Adesola; Okoh, Anthony I
Cocos nucifera belong to the family Aracaceae (palm Family). The English name is coconut and it is used extensively as medicinal remedies against infections such as urinary tract infections, gastro intestinal infections, skin and wound infections. The in vitro antibacterial (including anti-listerial and anti-vibrio) properties as well as the evaluation of the combination potentials of the plant extract with six front-line antibiotics were evaluated in this study using standard procedures. The in vitro anti-listerial properties of the crude aqueous and n-Hexane extract of the husk of Cocos nucifera were carried out against 37 Listeria isolates. Twenty-nine of the test organisms were susceptible to the aqueous extract while thirty were susceptible to the n-Hexane extract both at the screening concentration of 25 mg/ml. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.6 - 5.0 mg/ml. For the aqueous extract, average log reduction in viable cell count ranged between 0.32 Log10 and 4.8 Log10 CFU/ml after 8 hours interaction in 1 × MIC and 2 × MIC. For the n-Hexane extract, the log reduction ranged between 2.4 Log10 and 6.2 Log10 CFU/ml after 8 hours interaction in 1 × MIC and 2 × MIC. The time-kill characteristics of the two extracts suggest that at higher concentration (2 × MIC) and longer duration of interaction (8 hr), more bacteria were killed. In vitro anti-vibrio and antibacterial properties experiment revealed that of all the 45 vibrio and 25 bacteria strains that was tested, 37 were susceptible to the aqueous extract and 38 to the n-Hexane extract, while 17 were susceptible to the aqueous extract and 21 to the n-Hexane extract. Minimum Inhibitory Concentration (MIC) values for all the susceptible bacteria ranged between 0.3 - 5.0 mg/ml. viii The time kill studies revealed that for the aqueous extract, average log reduction in viable cell count in time kill assay ranged between 0.12 Log10 and 4.2 Log10 CFU/ml after 8 hr interaction at 1 × MIC and 2 × MIC. For the n-Hexane extract, the log reduction ranged between 0.56 Log10 and 6.4 Log10 CFU/ml after 8 hr interaction in 1 × MIC and 2 × MIC. In the test for the combination interactions, the checkerboard method revealed synergy of 67% and indifferent of 33%, while the time kill assay detected synergy in 72% and indifferent in 28% of the combinations tested. The synergy detected was not specific to any of the antibiotics or the Gram reaction of the bacteria, and no antagonism was detected. We conclude that the aqueous and n-Hexane extract of the husk of C. nucifera contains potential broad spectrum antibiotics resistance modulating compounds that could be relevant in the treatment of infections caused by these pathogens. In addition, the husk which is being discarded as agro waste will opens up a vista of opportunities for utilization for therapeutic purposes
Assessment of the flocculating efficiency of bioflocculant produced by bacillus sp. Aemreg4 isolated from Tyhume river, Eastern Cape, South Africa
(University of Fort Hare, 2016) Ntsangani, Nozipho; Mabinya, L V; Okoh, Anthony I
Bioflocculants are flocculating substances produced by microorganisms during growth and have recently received considerable attention from researchers; due to their biodegradability, non-toxicity and lack of secondary pollution from degradation intermediates. This study evaluated the efficiency of bioflocculant produced by Bacillus sp. AEMREG4 isolated from Tyhume River. The bacterial identification was through 16S rDNA sequencing; nucleotide sequences were deposited in GenBank as Bacillus sp. AEMREG4 with an Accession number KP406729. The optimum culture conditions for bioflocculant production were an inoculum size of 4% (v/v) and starch as well as yeast extract as sole carbon and nitrogen sources respectively. The addition of CaCl2 enhanced the flocculating activity, at a wide range of pH 4-10 and the highest flocculating activity was reached at an initial pH 8 (80%). A bioflocculant yield of 0.78 g was recovered from 1 L of culture broth. The optimum flocculating activity of 78% was reached at the lowest bioflocculant dosage of 0.1 mg/ml and the presence of divalent cations (Ca2+, Mn2+ and Mg2+) as well as a trivalent cation (Al3+) enhanced flocculating activity. The purified bioflocculant retained more than 70% flocculating activity when subjected to heating at 100 °C for 1 h and maximum flocculating activity of 83% was achieved at both acidic and basic pH values of 3 and 10 respectively. Chemical analysis showed that the bioflocculant is predominantly polysaccharide. The Fourier transform infrared (FTIR) spectrum revealed the presence of carboxyl, hydroxyl and methoxyl groups as the functional moieties and the scanning electron microscopy (SEM) imaging of the purified bioflocculant showed its morphological structure as rod-shaped which contributes to its high flocculating efficiency. The high flocculation activity displayed by this bioflocculant indicates its potential suitability for industrial application.Keywords: Bioflocculant, Bacillus sp. AEMREG4, flocculating activity.
Assessment of the flocculating efficiency of Bioflocculant produced by bacillus sp. Aemreg4 isolated from Tyhume river, Eastern Cape, South Africa.
(University of Fort Hare, 2016) Ntsangani, Nozipho
Bioflocculants are flocculating substances produced by microorganisms during growth and have recently received considerable attention from researchers; due to their biodegradability, non-toxicity and lack of secondary pollution from degradation intermediates. This study evaluated the efficiency of bioflocculant produced by Bacillus sp. AEMREG4 isolated from Tyhume River. The bacterial identification was through 16S rDNA sequencing; nucleotide sequences were deposited in GenBank as Bacillus sp. AEMREG4 with an Accession number KP406729. The optimum culture conditions for bioflocculant production were an inoculum size of 4% (v/v) and starch as well as yeast extract as sole carbon and nitrogen sources respectively. The addition of CaCl2 enhanced the flocculating activity, at a wide range of pH 4-10 and the highest flocculating activity was reached at an initial pH 8 (80%). A bioflocculant yield of 0.78 g was recovered from 1 L of culture broth. The optimum flocculating activity of 78% was reached at the lowest bioflocculant dosage of 0.1 mg/ml and the presence of divalent cations (Ca2+, Mn2+ and Mg2+) as well as a trivalent cation (Al3+) enhanced flocculating activity. The purified bioflocculant retained more than 70% flocculating activity when subjected to heating at 100 °C for 1 h and maximum flocculating activity of 83% was achieved at both acidic and basic pH values of 3 and 10 respectively. Chemical analysis showed that the bioflocculant is predominantly polysaccharide. The Fourier transform infrared (FTIR) spectrum revealed the presence of carboxyl, hydroxyl and methoxyl groups as the functional moieties and the scanning electron microscopy (SEM) imaging of the purified bioflocculant showed its morphological structure as rod-shaped which contributes to its high flocculating efficiency. The high flocculation activity displayed by this bioflocculant indicates its potential suitability for industrial application.
Assessment of the incidence of E.coli in Tyume and Buffalo rivers in the Eastern Cape Province of South Africa
(University of Fort Hare, 2013) Koba, Siziwe; Okoh, Anthony I
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
Assessment of the prevalence of faecal coliforms and Escherichia coli o157:h7 in the final effluents of two wastewater treatment plants in Amahlathi Local Municipality of Eastern Cape Province, South Africa
(University of Fort Hare, 2014) Ajibade, Adefisoye Martins; Green, E; Okoh, Anthony I
The production of final effluents that meet discharged requirements and guidelines remain a major challenge particularly in the developing world with the resultant problem of surface water pollution. This study assessed the physicochemical and microbiological qualities of two wastewater treatment works in the Eastern Cape Province of South Africa in terms of the prevalence of faecal coliforms and Escherichia coli O157:H7 over a five month period. All physicochemical and microbiological analyses were carried out using standard methods. Data were collected in triplicates and analysed statistically using IBM SPSS version 20.0. The ranges of some of the physicochemical parameters that complied with set guidelines include pH (6.7 – 7.6), TDS (107 – 171 mg/L), EC (168 – 266 μS/cm), Temperature (15 – 24oC), NO3- (0 – 8.2 mg/L), NO2- (0.14 – 0.71 mg/L) and PO4 (1.05 – 4.50 mg/L). Others including Turbidity (2.64 – 58.00 NTU), Free Cl (0.13 – 0.65 mg/L), DO (2.20 – 8.48 mg/L), BOD (0.13 – 6.85 mg/L) and COD (40 – 482 mg/L) did not comply with set guidelines. The microbiological parameters ranged 0 – 2.7 × 104 CFU/100 ml for FC and 0 – 9.3 × 103 for EHEC CFU/100 ml, an indication of non-compliance with set guidelines. Preliminary identification of 40 randomly selected presumptive enterohemorrhagic E. coli isolates by Gram’s staining and oxidase test shows 100% (all 40 selected isolates) to be Gram positive while 90% (36 randomly selected isolates) were oxidase negative. Statistical correlation between the physicochemical and the microbiological parameters were generally weak except in the case of free chlorine and DO where they showed inverse correlation with the microbiological parameters. The recovery of EHEC showed the inefficiency of the treatment processes to effectively inactivate the bacteria, and possibly other pathogenic bacteria that may be present in the treated wastewater. The assessment suggested the need for proper monitoring and a review of the treatment procedures used at these treatment works.
Assessment of the prevalence of virulent escherichia coli strains in the final effluents of wastewater treatment plants in the Eastern Cape Province of South Africa
(University of Fort Hare, 2010) Osode, Augustina Nwabuje
Escherichia coli (E. coli) is a common inhabitant of surface waters in the developed and eveloping 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 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 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.
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
(University of Fort Hare, 2010) Osode, Augustina Nwabuje; Okoh, Anthony I
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.
Assessment of the quality indices and prevalence of Escherichia coli pathotypes in selected rivers of Osun state, Southwestern Nigeria
(2015) Titilawo, Osuolale Yinka; ;
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.

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